Pubmed abstracts for Protein-Protein Interaction search result :


Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.67065283
It is thus apparent that Akt directly activates eNOS in endothelial cells . 0.67065283^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :1.8669635
Accordingly , we showed that early VEGF stimulation first leads to the Ca ( 2+ ) / calmodulin disruption of the caveolin eNOS complex and promotes the association between eNOS and hsp 90 . eNOS bound hsp 90 can then recruit VEGF activated ( phosphorylated ) Akt to the complex , which in turn can phosphorylate eNOS . 1.8669635^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.68874733
S1P activates the endothelial isoform of nitric oxide synthase ( eNOS ) , associated with eNOS phosphorylation at Ser 1179 , a site phosphorylated by protein kinase Akt . 0.68874733^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Recent studies show that intracellular signal protein kinase B ( PKB ) is involved in the regulation of cNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Myocardial hypoxia reoxygenation ( H R ) is associated with upregulation of inducible nitric oxide synthase ( iNOS ) , decrease in endothelial NOS ( eNOS ) , and increase in protein kinase B ( PKB ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In the penis , nitric oxide ( NO ) can be formed by both neuronal NO synthase and endothelial NOS ( eNOS ) . eNOS is activated by viscous drag / shear stress in blood vessels to produce NO continuously , a process mediated by the phosphatidylinositol 3 kinase ( PI3kinase ) / Akt pathway . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Western blot analysis showed that the extent of phosphorylation of Akt , an active form of Akt , was increased by cerivastatin while it was reduced by LY 294002 , suggesting an involvement of PI 3 kinase / Akt dependent activation of endothelial NOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Vanadate activates the phosphoinositide 3 kinase ( PI 3 K ) / Akt pathway , which is known to increase endothelial NOS ( eNOS ) activity by direct phosphorylation of Ser 1179 . ^^^ Treatment of BLMVEC with vanadate resulted in phosphorylation of both Akt and endothelial NOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Here we show that the serine / threonine protein kinase Akt ( protein kinase B ) can directly phosphorylate eNOS on serine 1179 and activate the enzyme , leading to NO production , whereas mutant eNOS ( S1179A ) is resistant to phosphorylation and activation by Akt . ^^^ Thus , eNOS is a newly described Akt substrate linking signal transduction by Akt to the release of the gaseous second messenger NO . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Here we demonstrate that the serine / threonine protein kinase Akt / PKB mediates the activation of eNOS , leading to increased NO production . ^^^ Inhibition of the phosphatidylinositol 3 OH kinase / Akt pathway or mutation of the Akt site on eNOS protein ( at serine 1177 ) attenuates the serine phosphorylation and prevents the activation of eNOS . ^^^ Thus , phosphorylation of eNOS by Akt represents a novel Ca2+ independent regulatory mechanism for activation of eNOS . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Because the sequence ( RIR ) TQpSFSLQER contains a consensus substrate site for protein kinase B ( PKB or Akt ) , we demonstrated that LY 294002 , an inhibitor of the upstream activator of PKB , phosphatidylinositol 3 kinase , inhibited flow induced eNOS phosphorylation by 97 % and NO production by 68 % . ^^^ Finally , PKB phosphorylated eNOS in vitro at the same site phosphorylated in the cell and increased eNOS enzymatic activity by 15 20 fold . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Inhibition of phosphatidylinositol 3 kinase ( PI3K ) , using wortmannin or a dominant negative mutant of its downstream target , Akt ( protein kinase B ) , prevented the maintained serine phosphorylation and activation of eNOS . ^^^ These data suggest that shear stress activates a pathway involving PI3K and the serine / threonine kinase Akt , which phosphorylates eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Bovine endothelial nitric oxide synthase ( eNOS ) is phosphorylated directly by the protein kinase Akt at serine 1179 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Using co immunoprecipitation experiments , we isolated eNOS in a complex with the MAP kinases extracellular signal regulated kinases 1 and 2 ( ERK1 / 2 ) as well as the protein kinases Raf 1 and Akt . ^^^ Within minutes of adding bradykinin to BAEC , the eNOS Raf 1 ERK Akt heteromeric complex dissociated , and it subsequently reassociated following more prolonged agonist stimulation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The protein kinase Akt activates the endothelial NO synthase ( eNOS ) by phosphorylation of Ser 1177 . ^^^ Therefore , we investigated the contribution of Akt mediated eNOS phosphorylation to VEGF induced EC migration . ^^^ Moreover , transfection of an Akt site phospho mimetic eNOS ( S1177D ) potently stimulated EC migration , whereas a non phosphorylatable mutant ( S1177A ) inhibited VEGF induced EC migration . ^^^ Our data indicate that eNOS activation via phosphorylation of Ser 1177 by Akt is necessary and sufficient for VEGF mediated EC migration . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The serine / threonine protein kinase Akt ( protein kinase B ) phosphorylates endothelial cell nitric oxide synthase ( eNOS ) and enhances its ability to generate nitric oxide ( NO ) . ^^^ L NAME , an eNOS inhibitor , blocked myr Akt mediated vasodilatation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Accordingly , simvastatin enhanced phosphorylation of the endogenous Akt substrate endothelial nitric oxide synthase ( eNOS ) , inhibited apoptosis and accelerated vascular structure formation in vitro in an Akt dependent manner . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Either treatment with wortmannin or exogenous expression of a dominant negative Akt in TRLECs decreased the 17beta E 2 induced eNOS activation . ^^^ Moreover , 17beta E 2 induced Akt activation actually enhances the phosphorylation of eNOS . 17beta E 2 induced Akt activation was dependent on both extracellular and intracellular Ca ( 2+ ) . ^^^ Our findings suggest that 17beta E 2 induced eNOS activation through an Akt dependent mechanism , which is mediated by ERalpha via a nongenomic mechanism . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Membrane estrogen receptor engagement activates endothelial nitric oxide synthase via the PI 3 kinase Akt pathway in human endothelial cells . 17beta Estradiol ( E ( 2 ) ) is a rapid activator of endothelial nitric oxide synthase ( eNOS ) . ^^^ Here we demonstrate for the first time , to our knowledge , that E ( 2 ) rapidly induces phosphorylation and activation of eNOS through the phosphatidylinositol 3 ( PI 3 ) kinase Akt pathway . ^^^ As has been shown for vascular endothelial growth factor , eNOS is an E ( 2 ) activated Akt substrate , demonstrated by rapid eNOS phosphorylation on serine 1177 , a critical residue for eNOS activation and enhanced sensitivity to resting cellular Ca ( 2+ ) levels . ^^^ The membrane impermeant E ( 2 ) BSA conjugate , shown to bind endothelial cell membrane sites , also induced rapid Akt and consequent eNOS phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Production of nitric oxide ( NO ) in endothelial cells is regulated by direct interactions of endothelial nitric oxide synthase ( eNOS ) with effector proteins such as Ca2+ calmodulin , by posttranslational modifications such as phosphorylation via protein kinase B , and by translocation of the enzyme from the plasma membrane caveolae to intracellular compartments . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
S1P treatment activates Akt , a protein kinase implicated in phosphorylation of eNOS . ^^^ S1P treatment of BAEC leads to eNOS phosphorylation at Ser ( 1179 ) , a residue phosphorylated by Akt ; an eNOS mutant in which this Akt phosphorylation site is inactivated shows attenuated S1P induced eNOS activation . ^^^ S1P induced activation both of Akt and of eNOS is inhibited by pertussis toxin , by the phosphoinositide 3 kinase inhibitor wortmannin , and by the intracellular calcium chelator BAPTA ( 1 , 2 bis ( aminophenoxy ) ethane N , N , N ' , N ' tetraacetic acid ) . ^^^ By contrast to S1P , activation of G protein coupled bradykinin B 2 receptors neither activates kinase Akt nor promotes Ser ( 1179 ) eNOS phosphorylation despite robustly activating eNOS enzyme activity . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Here we show that SPP signals through the EDG 1 receptor to the heterotrimeric G protein G ( 1 ) , leading to activation of the serine / threonine kinase Akt and phosphorylation of the Akt substrate , endothelial nitric oxide synthase ( eNOS ) . ^^^ SPP also stimulates eNOS phosphorylation and NO release and these effects are also attenuated by inhibition of G ( 1 ) signaling , PI 3 kinase , and Akt . ^^^ Thus , SPP signals through G ( 1 ) and PI 3 kinase leading to Akt activation and eNOS phosphorylation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Both vascular endothelial growth factor and FSS activate endothelial protein kinase B ( PKB ) by way of incompletely understood pathway ( s ) , and , in turn , PKB phosphorylates eNOS at Ser 1179 , causing its activation . ^^^ In this study , we found that either FSS or insulin stimulated insulin receptor substrate 1 ( IRS 1 ) tyrosine and serine phosphorylation and increased IRS 1 associated phosphatidylinositol 3 kinase activity , phosphorylation of PKB Ser 473 , phosphorylation of eNOS Ser 1179 , and NO production . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Stimulating ECs with vascular endothelial growth factor ( VEGF ) leads to the activation of Akt / protein kinase B , which in turn activates endothelial nitric oxide synthase ( eNOS ) by phosphorylation on serine 1177 . ^^^ Therefore , we investigated whether oxLDL affects EC migration by an inhibitory effect on the Akt / eNOS pathway . ^^^ Overexpression of a constitutively active Akt construct ( Akt T308D / S473D ) or a phosphomimetic eNOS construct ( eNOS S1177D ) almost completely reversed the inhibitory effect of oxLDL on VEGF induced EC migration and NO generation . ^^^ CONCLUSIONS : Our data indicate that oxLDL induced dephosphorylation of Akt , followed by impaired eNOS activation , reduces the intracellular level of NO and thereby inhibits VEGF induced EC migration . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Endothelial nitric oxide synthase ( eNOS ) is phosphorylated at Ser 1179 ( bovine sequence ) by Akt after growth factor or shear stress stimulation of endothelial cells , resulting in increased eNOS activity . ^^^ We investigated whether bradykinin ( BK ) stimulation of bovine aortic endothelial cells ( BAECs ) regulates eNOS through Akt activation and Ser 1179 or Thr 497 phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Insulin stimulated activation of eNOS is independent of Ca2+ but requires phosphorylation by Akt at Ser ( 1179 ) . ^^^ However , cells expressing the eNOS S1179A mutant ( disrupted Akt phosphorylation site ) did not produce detectable NO in response to insulin , whereas the response to LPA was similar to that observed in cells expressing wild type eNOS . ^^^ Interestingly , platelet derived growth factor treatment of cells activated Akt but not eNOS . ^^^ We conclude that insulin regulates eNOS activity using a Ca ( 2+ ) independent mechanism requiring phosphorylation of eNOS by Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In endothelial cells , the VEGF / Flk 1 / KDR signal system is a very important generator of nitric oxide ( NO ) through the activation of its downstream effectors phosphatidylinositol 3 OH kinase ( PI 3 K ) , Akt kinase and endothelial NO synthase ( eNOS ) . ^^^ By using the OCI / AML 2 cell line , which expresses VEGF receptor 2 , ie Flk 1 / KDR , eNOS and VEGF , as analyzed by flow cytometry , and produces VEGF into growth medium , as analyzed by ELISA , we showed that the Akt kinase and NOS activities in these cells were decreased by the inhibitors of VEGF , Flk 1 / KDR and PI 3 K , and NOS activity also by the direct inhibitor of NOS . ^^^ We conclude that autocrine VEGF through Flk 1 / KDR , by activating eNOS to produce NO through PI 3 K / Akt kinase , maintains clonogenic cell growth in the OCI / AML 2 cell line . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
A recent paper has expanded the regulation of the enzyme to the realm of sphingolipid signaling , specifically implicating that sphingosine 1 phosphate , endothelial differentiation gene ( Edg ) receptors and Akt kinase induce a signal transduction pathway via phosphorylation of a serine residue in eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The effect of estrogen on cardiovascular disease is mainly dependent on direct actions on the vascular wall involving activation of endothelial nitric oxide synthase ( eNOS ) via Akt and extracellular signal regulated protein kinase ( ERK ) cascades . ^^^ Either exogenous expression of a dominant negative Akt or pretreatment of TRLECs with PD 98059 decreased the raloxifene induced eNOS phosphorylation . ^^^ Moreover , raloxifene stimulated the activation of Akt , ERK , and eNOS in Chinese hamster ovary cells expressing estrogen receptor alpha but not Chinese hamster ovary cells expressing estrogen receptor beta . ^^^ Our findings suggest that raloxifene induced eNOS phosphorylation is mediated by estrogen receptor alpha via a nongenomic mechanism and is differentially mediated by Akt and ERK dependent cascades . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Using geldanamycin , an inhibitor of hsp 90 function , and overexpression of recombinant hsp 90 , we documented that the statin induced phosphorylation of eNOS on Ser 1177 was directly dependent on the ability of hsp 90 to recruit Akt in the eNOS complex . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Caveolin 1 , 90 kDa heat shock protein , ERK 1 / 2 , and Akt , all known and proposed regulators of eNOS activity , were detected throughout the ovine adrenal cortex . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Endothelial nitric oxide synthase ( eNOS ) is activated by phosphorylation of serine 1177 by the protein kinase Akt / PKB . ^^^ In contrast , when myc tagged human eNOS carried a mutation at the Akt phosphorylation site ( Ser 1177 ) , O linked N acetylglucosamine modification was unchanged by hyperglycemia and phospho eNOS was undetectable . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Because eNOS is activated by Akt dependent phosphorylation to produce nitric oxide ( NO ) , we determined the subcellular distribution of eNOS phosphorylated on serine 1179 using a variety of methodologies . ^^^ Co transfection of eNOS with Akt and stimulation of endothelial cells with vascular endothelial growth factor ( VEGF ) increased the ratio of P eNOS to total eNOS but did not change the relative intracellular distribution between these domains . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Akt has been believed to regulate shear dependent production of NO ( * ) by directly phosphorylating endothelial nitric oxide synthase ( eNOS ) at the Ser ( 1179 ) residue ( eNOS S ( 1179 ) ) , but a critical evaluation using specific inhibitors or dominant negative mutants ( Akt ( AA ) or Akt ( AAA ) ) has not been reported . ^^^ Here , we show that shear dependent phosphorylation of eNOS S ( 1179 ) is mediated by an Akt independent , but a PKA dependent , mechanism . ^^^ Expression of Akt ( AA ) or Akt ( AAA ) in BAEC by using recombinant adenoviral constructs inhibited phosphorylation of eNOS S ( 1179 ) if cells were stimulated by vascular endothelial growth factor ( VEGF ) , but not by shear stress . ^^^ In contrast , H 89 did not inhibit phosphorylation of eNOS S ( 1179 ) induced by expressing a constitutively active Akt mutant ( Akt ( Myr ) ) in BAEC , showing that the inhibitor did not affect the Akt pathway . 8 Bromo cAMP alone phosphorylated eNOS S ( 1179 ) within 5 min without activating Akt , in an H 89 sensitive manner . ^^^ Collectively , these results demonstrate that shear stimulates phosphorylation of eNOS S ( 1179 ) in a PKA dependent , but Aktindependent manner , whereas the NO ( * ) production is regulated by the mechanisms dependent on both PKA and Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Our results show that NO from eNOS inhibiting redox sensitive MCP 1 expression is mediated via Rac dependent NADPH oxidase by reducing ROS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Concomitant with eNOS activation , H ( 2 ) O ( 2 ) also activated Akt , increased eNOS phosphorylation at Ser 1177 , and decreased eNOS phosphorylation at Thr 495 . ^^^ We found that Akt activation , eNOS Ser 1177 phosphorylation , and eNOS activation by H ( 2 ) O ( 2 ) were calcium dependent , whereas eNOS dephosphorylation at Thr 495 was not , suggesting a branch point in the signaling cascade downstream from PI 3 K . ^^^ Consistent with this , overexpression of a dominant negative isoform of Akt inhibited H ( 2 ) O ( 2 ) induced phosphorylation of eNOS at Ser 1177 but not dephosphorylation of eNOS at Thr 495 . ^^^ In turn , PI 3 K mediates eNOS Ser 1177 phosphorylation via a calcium and Akt dependent pathway , whereas eNOS Thr 495 dephosphorylation does not involve calcium or Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In conclusion , leptin induces NO production by activating a PI 3 kinase independent Akt eNOS phosphorylation pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
S1P activated the phosphatidylinositol 3 kinase ( PI3K ) / Akt / endothelial NO synthase ( eNOS ) pathway in ECs , since S1P stimulated eNOS phosphorylation and NO production were blocked by inhibition of activities of PI3K and Akt . ^^^ S1P stimulated EC migration and tube formation on Matrigel , which processes were significantly decreased by inhibition of activities of PI3K , Akt , or eNOS , whereas treatment with LY 294002 , a PI3K inhibitor , but not L NAME , inhibited EC viability and proliferation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Western blot analysis was performed to determine the activation of p 38 , AKT , eNOS . ^^^ RESULTS : Arsenite induced the activation of AKT at both Ser 473 and Thr 308 , and its downstream effector eNOS in cultured human keratinocytes . ^^^ PI 3 kinase inhibitors , Wortmannin and LY 294002 inhibited arsenite induced phosphorylation of AKT and eNOS but had no effect on phosphorylation of p 38 . ^^^ Interestingly , however , SB 203580 , a known p 38 inhibitor , completely inhibited arsenite induced phosphorylation of AKT and eNOS . ^^^ CONCLUSIONS : Collectively , our data indicate that arsenite induces activation of AKT and eNOS , via PI 3 kinase and p 38 pathway , likely bypassing the activation of EGF receptor in cultured human keratinocytes . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Importantly , we have extended these studies to examine the mechanism whereby HDL binding to SR BI stimulates eNOS . eNOS can be stimulated by an increase in intracellular calcium , by phosphorylation by Akt kinase , or by an increase in intracellular ceramide . ^^^ We conclude that HDL binding to SR BI stimulates eNOS by increasing intracellular ceramide levels and is independent of an increase in intracellular calcium or Akt kinase phosphorylation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Activation of eNOS in rat portal hypertensive gastric mucosa is mediated by TNF alpha via the PI 3 kinase Akt signaling pathway . ^^^ In PHT gastric mucosa we studied ( 1 ) eNOS phosphorylation ( at serine 1177 ) required for its activation ; ( 2 ) association of the phosphatidylinositol 3 kinase ( PI 3 kinase ) , and its downstream effector Akt , with eNOS ; and , ( 3 ) whether TNF alpha neutralization affects eNOS phosphorylation and PI 3 kinase Akt activation . ^^^ PHT gastric mucosa has significantly increased ( 1 ) eNOS phosphorylation at serine 1177 by 90 % ( P < . 01 ) ; ( 2 ) membrane translocation ( P < . 05 ) and phosphorylation ( P < . 05 ) of p 85 ( regulatory subunit of PI 3 kinase ) by 61 % and 85 % , respectively ; ( 3 ) phosphorylation ( P < . 01 ) and activity ( P < . 01 ) of Akt by 40 % and 52 % , respectively ; and ( 4 ) binding of Akt to eNOS by as much as 410 % ( P < . 001 ) . ^^^ Neutralizing anti TNF alpha antibody significantly reduced p 85 phosphorylation , phosphorylation and activity of Akt , and eNOS phosphorylation in PHT gastric mucosa to normal levels . ^^^ In conclusion , these findings show that in PHT gastric mucosa , TNF alpha stimulates eNOS phosphorylation at serine 1177 ( required for its activation ) via the PI 3 kinase Akt signal transduction pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
BACKGROUND : Recent evidence from cultured endothelial cell studies suggests that phosphorylation of endothelial nitric oxide synthase ( eNOS ) through the PI 3 kinase Akt pathway increases NO production . ^^^ Treatment with insulin resulted in 2 . 6 fold and 4 . 3 fold increases in Akt and eNOS phosphorylation and a significant increase in NO production in ischemic / reperfused myocardial tissue . ^^^ Phosphorylation of Akt and eNOS and increase of NO production by insulin were completely blocked by wortmannin , a PI 3 kinase inhibitor . ^^^ Pretreatment with L NAME , a nonselective NOS inhibitor , had no effect on Akt and eNOS phosphorylation but significantly reduced NO production . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The dose dependent promigratory and proangiogenic effects of atorvastatin on mature endothelial cells are correlated with the activation of the phosphatidylinositol 3 kinase Akt pathway , as determined by the phosphorylation of Akt and endothelial NO synthase ( eNOS ) at Ser 1177 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
BACKGROUND AND PURPOSE : Bovine endothelial nitric oxide synthase ( eNOS ) is phosphorylated directly by the protein kinase Akt at serine 1179 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We now report that LPA treatment of bovine aortic endothelial cells ( BAEC ) activates eNOS enzyme activity in a pathway that involves phosphorylation of eNOS on serine 1179 by protein kinase Akt . ^^^ In contrast to the cellular responses elicited by S1P in COS 7 cells , LPA can stimulate the activation of eNOS and Akt independently of EDG 1 receptor transfection . ^^^ LPA stimulated enzyme activation was significantly attenuated in an eNOS mutant lacking the site that is phosphorylated by kinase Akt ( eNOS S1179A ) . ^^^ Taken together , these results indicate that LPA potently activates eNOS in BAEC in a pathway distinct from the EDG 1 receptor , but mediated by a similar receptor mediated pathway dependent on pertussis toxin sensitive G proteins and involving activation of the PI 3 K / Akt pathway . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Because the activation of eNOS by VEGF is associated with the Akt dependent phosphorylation of eNOS at Ser 1177 , we investigated whether endostatin interferes with phosphorylation of eNOS . ^^^ Endostatin reduced VEGF induced phosphorylation of eNOS at Ser 1177 , whereas Akt phosphorylation was not affected . ^^^ Finally , a non dephosphorylatable constitutive active eNOS construct ( S1177D ) , but not constitutive active Akt , abolished the inhibitory effect of endostatin on EC migration . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The catalytic activity of eNOS is augmented by phosphorylation of a C terminal serine residue ( Ser 1177 of human eNOS ) through the phosphatidyl 3 kinase ( PI 3K ) / Akt pathway . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
However , ONOO ( ) strongly inhibited the phosphorylation and activity of Akt , which is known to phosphorylate eNOS Ser ( 1179 ) . ^^^ Moreover , expression of an Akt dominant negative mutant did not prevent ONOO ( ) enhanced eNOS Ser ( 1179 ) phosphorylation . ^^^ We conclude that ONOO ( ) inhibits Akt and increases AMPK dependent Ser ( 1179 ) phosphorylation of eNOS resulting in enhanced O ( 2 ) ( ) release . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Both in vivo and in vitro phosphorylation of endothelial nitric oxide synthase ( eNOS ) by Akt were reduced by hyperglycemia and hexosamine activation . ^^^ CONCLUSIONS : Our data show that hyperglycemia , through the hexosamine pathway , impairs activation of the IR / IRS / PI3 K / Akt pathway , resulting in deregulation of eNOS activity . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Vasodilator actions of insulin are mediated by signaling pathways involving phosphatidylinositol 3 kinase ( PI 3 kinase ) and Akt that lead to activation of endothelial nitric oxide synthase ( eNOS ) in endothelium . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Purified protein kinase A ( PKA ) phosphorylates both sites in purified eNOS , whereas purified Akt phosphorylates only Ser ( 617 ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Furthermore , eNOS Ser ( 1177 ) phosphorylation in both conditions significantly decreased without affecting Akt phosphorylation in the hypoxic artery . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The wild type eNOS underwent agonist induced phosphorylation at serine 1179 ( a putative site for phosphorylation by kinase Akt ) , but phosphorylation of the myr ( ) eNOS at this residue was nearly abrogated ; the palm ( ) eNOS exhibited an intermediate phenotype . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Although chronic hypoxia decreased phosphorylated eNOS ( S 1177 ) levels by approximately 4 to 5 fold and total Akt and phosphorylated Akt by 4 and 5 fold , it also increased hsp 90 association with eNOS by more than 3 fold . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Transactivation of vascular endothelial growth factor ( VEGF ) receptor Flk 1 / KDR is involved in sphingosine 1 phosphate stimulated phosphorylation of Akt and endothelial nitric oxide synthase ( eNOS ) . ^^^ S1P stimulation of eNOS phosphorylation was shown to involve G ( 1 ) protein , phosphoinositide 3 kinase , and Akt . ^^^ VEGF also activates eNOS through Flk 1 / KDR , phosphoinositide 3 kinase , and Akt , which suggested that S1P and VEGF may share upstream signaling mediators . ^^^ S1P mediated phosphorylation of Flk 1 / KDR , Akt , and eNOS were all inhibited by VEGF receptor tyrosine kinase inhibitors and by antisense Flk 1 / KDR oligonucleotides . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We found no evidence for activation of eNOS via the serine / threonine kinase Akt / PKB ( protein kinase B ) in adenosine stimulated cells . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In contrast , multiple site disruption of a pathway leading to eNOS activation via the serine / threonine kinase Akt was ineffective . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Activation of the protein kinase Akt / PKB mediates VEGF dependent endothelial cell survival and eNOS activation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The effect of chronic hyperglycemia on endothelial NO synthase ( eNOS ) activity and expression , glycogen synthase ( GS ) activity , extracellular signal regulated kinase ( ERK 1 , 2 ) , p 38 , Akt expression , and Cu / Zn superoxide dismutse ( SOD 1 ) activity and expression were determined . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This interaction leads to activation of PI3K signaling cascade to Ser / Thr kinase Akt , which mediates several PI3K dependent intracellular effects , including endothelial isoform of NO synthase ( eNOS ) phosphorylation and activation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Because activation of two signaling pathways , the phosphatidylinositol 3 kinase ( PI3K ) > Akt > eNOS and the ERK1 / 2 MAPK pathway , is known to be involved in cell migration , we used the pharmacological inhibitors wortmannin and PD 98059 to determine if chemotactic signaling by leptin involves Akt or ERK1 / 2 , respectively . ^^^ Both PPARgamma ligands inhibited leptin stimulated Akt and eNOS phosphorylation , but neither attenuated ERK 1 / 2 activation in response to leptin . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Src kinase mediates phosphatidylinositol 3 kinase / Akt dependent rapid endothelial nitric oxide synthase activation by estrogen . 17beta Estradiol activates endothelial nitric oxide synthase ( eNOS ) , enhancing nitric oxide ( NO ) release from endothelial cells via the phosphatidylinositol 3 kinase ( PI 3 kinase ) / Akt pathway . ^^^ This complex formation results in the successive activation of PI 3 kinase , Akt , and eNOS with consequent enhanced NO release , implicating c Src as a critical upstream regulator of the estrogen stimulated PI 3 kinase / Akt / eNOS pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Using adenovirus mediated gene transfer in human umbilical vein endothelial cells ( HUVECs ) , we show here that both active RhoA and ROCK not only downregulate eNOS gene expression as reported previously but also inhibit eNOS phosphorylation at Ser 1177 and cellular NO production with concomitant suppression of PKB activation . ^^^ Moreover , coexpression of a constitutive active form of PKB restores the phosphorylation but not gene expression of eNOS in the presence of active RhoA . ^^^ Expression of the active PKB reverses eNOS phosphorylation but has no effect on downregulation of eNOS expression induced by thrombin . ^^^ Taken together , these data demonstrate that Rho / ROCK pathway negatively regulates eNOS phosphorylation through inhibition of PKB , whereas it downregulates eNOS expression independent of PKB . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This Akt mediated increase in vascular permeability was inhibited by the eNOS inhibitor L NAME . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
To determine the mechanism by which GR activated eNOS , we measured the effect of corticosteroids on PI3K and the protein kinase Akt . ^^^ These findings indicate that non nuclear GR rapidly activates eNOS through the PI3K / Akt pathway and suggest that this mechanism mediates the acute neuroprotective effects of corticosteroids through augmentation of CBF . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We surgically induced hindlimb ischemia in transgenic mice overexpressing endothelial NO synthase in the endothelium ( eNOS Tg ) and studied neocapillary formation , ischemia induced vascular endothelial growth factor ( VEGF ) expression , cGMP accumulation , and Akt / PKB signaling . ^^^ Basal and VEGF mediated Akt phosphorylation in aortas was similar between eNOS Tg and wild type mice . ^^^ Our study demonstrates that overexpression of eNOS protein causes a marked increase in neocapillary formation in response to tissue ischemia without affecting ischemia induced VEGF expression or VEGF mediated Akt phosphorylation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
A variety of stimuli regulate eNOS activity through signaling pathways involving Akt kinase and / or mitogen activated protein ( MAP ) kinase . ^^^ HDL ( 10 50 microg / ml , 20 min ) caused eNOS phosphorylation at Ser 1179 , and dominant negative Akt inhibited both HDL mediated phosphorylation and activation of the enzyme . ^^^ In addition , HDL activated MAP kinase through PI 3 kinase , and mitogen activated protein kinase / extracellular signal regulated kinase kinase inhibition fully attenuated eNOS stimulation by HDL without affecting Akt or eNOS Ser 1179 phosphorylation . ^^^ These results indicate that HDL stimulates eNOS through common upstream , Src mediated signaling , which leads to parallel activation of Akt and MAP kinases and their resultant independent modulation of the enzyme . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Recent investigations have shown that estrogen ' s rapid stimulatory action on eNOS is mediated by the activation of phosphatidylinositol 3 kinase ( PI 3 K ) and protein kinase B ( PKB ) / Akt pathway among other signaling systems . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Phosphorylation of endothelial nitric oxide synthase ( eNOS ) at serine 1179 was observed > 10 min after the addition of H ( 2 ) O ( 2 ) , and this was prevented by wortmannin but not by PD 98059 . c Src family tyrosine kinase ( s ) was found to be upstream of H ( 2 ) O ( 2 ) dependent Akt and eNOS serine 1179 phosphorylation and subsequent NO * production . ^^^ In summary , H ( 2 ) O ( 2 ) causes endothelial NO * release mediated by cooperative effects between PI 3 kinase / Akt dependent eNOS serine 1179 phosphorylation and activation of MEK / ERK1 / 2 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Phosphorylation of Akt and eNOS was evident in sheared fetal but not adult PAEC . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Wall stiffness suppresses Akt / eNOS and cytoprotection in pulse perfused endothelium . ^^^ Increased steady shear stress stimulates nitric oxide synthase ( eNOS ) in part by Akt dependent phosphorylation . ^^^ Here , we tested the hypothesis that reduced wall distensibility alters PP induced eNOS / Akt mechano signaling . ^^^ In compliant tubes , PP doubled Akt phosphorylation above nonpulsatile flow levels , whereas P Akt declined to static levels from PP in stiffer tubes . eNOS phosphorylation ( S 1179 ) similarly increased with PP in compliant tubes but was nearly undetectable with increased PP in stiffer tubes . ^^^ These data provide novel evidence that wall compliance can directionally mediate endothelial Akt / eNOS phosphorylation and mechano signaling . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Western blot analysis revealed that atorvastatin resulted in rapid activation of the PI3K / Akt signaling cascade ( within 5 min ) and that both Akt and eNOS phosphorylation were significantly increased by 4 . 1 fold and 2 . 9 fold , respectively ( p < 0 . 01 ) . ^^^ CONCLUSIONS : Atorvastatin attenuates lethal reperfusion induced injury in a manner that is reliant on PI3K and Akt activity and the presence and activity of eNOS . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We examined the influence of individual serine phosphorylation sites in endothelial nitric oxide synthase ( eNOS ) on basal and stimulated NO release , cooperative phosphorylation , and co association with hsp 90 and Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Abeta 42 generation is toxic to endothelial cells and inhibits eNOS function through an Akt / GSK 3beta signaling dependent mechanism . ^^^ Since the serine threonine kinase Akt is crucial to both neuronal and endothelial cell survival as well as eNOS activation , we investigated the effects of Abeta expression on Akt signaling in cultured endothelial cells . ^^^ Substrates downstream of Akt action , GSK 3beta and eNOS , are underphosphorylated in the presence of Abeta . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Bradykinin limits infarction when administered as an adjunct to reperfusion in mouse heart : the role of PI3K , Akt and eNOS . ^^^ Attenuation of reperfusion injury by growth factors has recently been linked to recruitment of phosphatidylinositol 3 kinase ( PI3K ) and protein kinase B ( Akt ) , a pathway also linked to the phosphorylation of eNOS by bradykinin . ^^^ Using western blot analysis , we show that bradykinin administration results in rapid , robust phosphorylation of both Akt and eNOS , greater than that seen in control hearts upon reperfusion ( Akt / eNOS phosphorylation : 68 + / 7 / 122 + / 29 AU versus 32 + / 5 / 47 + / 10 AU respectively , P < 0 . 01 ) . ^^^ This pattern of Akt phosphorylation was mimicked in the absence of eNOS , whereas Akt phosphorylation was inhibited by wortmannin . ^^^ Therefore , we demonstrate that exogenous bradykinin , administered at reperfusion , limits infarct size with concomitant rapid phosphorylation of Akt and eNOS , and that this protection is dependent upon the presence of eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Furthermore , statins may also increase eNOS activity via post translational activation of the phosphatidylinositol 3 kinase / protein kinase Akt ( PI 3 K / Akt ) pathway and / or through an interaction with the molecular chaperone heat shock protein 90 ( HSP 90 ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Recent studies have indicated that insulin activates endothelial nitric oxide synthase ( eNOS ) by protein kinase B ( PKB ) mediated phosphorylation at Ser 1177 in endothelial cells . ^^^ This was accompanied by reduced expression of IRS 2 and attenuated insulin stimulated recruitment of PI3K to IRS 1 and IRS 2 , yet insulin stimulated PKB activity and phosphorylation of eNOS at Ser 1177 were unaffected . ^^^ These data suggest that high glucose specifically inhibits insulin stimulated NO synthesis and down regulates some aspects of insulin signaling , including the CAP Cbl signaling pathway , yet this is not a result of reduced PKB mediated eNOS phosphorylation at Ser 1177 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Activation of endothelial nitric oxide synthase ( eNOS ) has been shown to occur through various pathways involving increases in the cytosolic Ca ( 2+ ) concentration , activation of the phosphatidylinositol 3 ' kinase / Akt pathway , as well as regulation by other kinases and by protein protein interactions . ^^^ We report here that eNOS activation in response to TNF alpha correlated with phosphorylation of Akt at Ser 473 and of eNOS itself at Ser 1179 . ^^^ Akt and eNOS phosphorylation , as well as eNOS activation , were blocked by inhibitors of both phosphatidylinositol 3 ' kinase and neutral sphingomyelinase . ^^^ Taken together , these results indicate that eNOS activation by TNF alpha occurs through sequential activation of neutral sphingomyelinase and of the phosphatidylinositol 3 ' kinase / Akt pathway . ^^^ The time course of eNOS activation induced through this pathway was markedly different from that triggered by ATP and epidermal growth factor , which activate eNOS through an increase in intracellular Ca ( 2+ ) concentration and through a sphingomyelinase independent stimulation of the phosphatidylinositol 3 ' kinase / Akt pathway , respectively . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Studies on endothelial cell cultures further revealed that VEGF stimulated phosphorylation of VEGF receptor 2 ( VEGFR 2 ) , leading to activation of Rac as well as increased phosphorylation of phospholipase Cgamma ( PLCgamma ) , protein kinase B ( Akt ) , endothelial nitric oxide synthase ( eNOS ) , and extracellular regulated kinase ( Erk1 / 2 ) . ^^^ We further found that phosphatidylinositol 3 OH kinase ( PI3K ) acted upstream of Rac and Akt eNOS in VEGF / VEGFR 2 signaling . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Activation of eNOS by VEGF involves several pathways including Akt / PKB , Ca ( 2+ ) / calmodulin , and protein kinase C . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Employing rat aortic EC ( RAEC ) , the effects of acute ( 20 and 30 minutes ) and prolonged ( 4 hours ) stimulation with 100 nmol / L IGF 1 and 1 nmol / L E 2 ( alone or in combination ) were assessed with respect to protein levels and enzymatic activities for phosphatidyl inositol 3 kinase ( PI3K ) and serine / threonine kinase Akt ( Akt ) , enzymes involved in eNOS activation . ^^^ The results suggest that IGF 1 and E 2 may interact through PI3K / Akt related pathways to increase eNOS activity . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In addition , eNOS activity and phosphorylation were abolished by pretreatment of the cells with an intracellular Ca ( 2+ ) chelator , bis ( o aminophenoxy ) ethane N , N , N ' , N ' tetra acetic acid tetrakis ( acetoxymethyl ester ) ( BAPTA / AM ) , with a suppression of Akt phosphorylation . ^^^ These results suggest that HGF stimulates eNOS activity by a PI3K / Akt dependent phosphorylation in a Ca ( 2+ ) sensitive manner in vascular endothelial cells . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Ang 1 has been shown to prevent EC apoptosis through activation of PI 3 kinase / Akt , a pathway that is also known to activate endothelium nitric oxide synthase ( eNOS ) . ^^^ Therefore , we hypothesized that the angiogenic effects of Ang 1 would also be dependent on the PI 3 kinase / Akt pathway , possibly mediated by increased eNOS activity and NO release . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In addition , G6PD deficient cells demonstrated decreased tyrosine phosphorylation of the VEGF receptor Flk 1 / KDR , Akt , and eNOS compared with cells with normal G6PD activity , whereas overexpression of G6PD enhanced phosphorylation of Flk 1 / KDR , Akt , and eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Expression of dominant negative AMPK attenuated AICAR stimulated AMPK activity , eNOS Ser 1177 phosphorylation and NO production and was without effect on AICAR stimulated protein kinase B Ser 473 phosphorylation or NO production stimulated by insulin or A 23187 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Synergistic activation of endothelial nitric oxide synthase ( eNOS ) by HSP 90 and Akt : calcium independent eNOS activation involves formation of an HSP 90 Akt CaM bound eNOS complex . ^^^ Here we extend these studies to explore interactions between HSP 90 , Akt , and eNOS . ^^^ In studies with purified proteins , HSP 90 increased the initial rate and maximal extent of Akt mediated eNOS phosphorylation and activation at low Ca2+ levels . ^^^ Akt was not observed in the eNOS complex in the absence of HSP 90 , but both active and inactive Akt associated with eNOS in the presence of HSP 90 . ^^^ Direct binding of Akt to HSP 90 was observed even in the absence of eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The protein kinase Akt is involved in embryonic vascular development and neoangiogenesis as well as in several endothelial cell functions , including activation of endothelial NO synthase ( eNOS ) and promotion of endothelial cell survival . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This acute effect of BK on both increases was blocked only by treatment of protein kinase A inhibitor H 89 , but not by the inhibitors of calmodulin dependent kinase 2 and protein kinase B , suggesting that the rapid increase in NO production by BK is mediated by the PKA dependent phosphorylation of eNOS Ser ( 1179 ) . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Both HDL and apoA 1 activated mitogen activated protein kinase and phosphatidylinositol 3 kinase ( PI3K ) Akt pathways in human arterial endothelial cells , and inhibition of either of these pathways by specific pharmacologic inhibitors abolished the effect of HDL on eNOS . ^^^ CONCLUSIONS : We demonstrate that HDL activates both extracellular signal regulated kinase 1 / 2 ( ERK1 / 2 ) and Akt , resulting in enhanced eNOS protein stability and subsequent accumulation of eNOS protein . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
METHODS : First , we determined the expression of eNOS in 3T3 L 1 adipocytes , and then these cells were treated with the NO donor sodium nitroprusside ( SNP ) and / or insulin , and glucose uptake and phosphorylation of insulin receptor substrate ( IRS ) 1 and Akt were evaluated . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The aim of the present study was to investigate the effects of exercise training on the endothelial function in relation to the expression of eNOS and Akt dependent eNOS phosphorylation in the left internal mammary artery ( LIMA ) of patients with stable CAD . ^^^ In LIMA tissue sampled during bypass surgery , eNOS expression and content of pospho eNOS Ser 1177 , Akt , and phospho Akt were determined by Western blot and quantitative reverse transcriptase polymerase chain reaction . ^^^ A linear correlation was confirmed between Akt phosphorylation and phospho eNOS levels ( R=0 . 80 , P < 0 . 05 ) and between phospho eNOS and Delta APV ( R=0 . 59 , P < 0 . 05 ) . ^^^ The change in acetylcholine induced vasodilatation was closely related to a shear stress induced / Akt dependent phosphorylation of eNOS on Ser1177 . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
After 14 days , cell ingrowth in the Matrigel plug was increased by 2 . 0 and 2 . 5 fold in rhVEGF B 167 treated and rhVEGF A 165 treated mice , respectively ( P < 0 . 01 ) , in association with a raise in phospho Akt / Akt ( 1 . 8 fold , P < 0 . 01 ) and endothelial NO synthase ( eNOS ) ( 1 . 80 and 1 . 60 fold , respectively ; P < 0 . 05 ) protein levels measured by Western blot . ^^^ Such an effect was associated with an upregulation of phospho Akt / Akt and eNOS protein levels in the ischemic legs and was hampered by treatment with anti VEGFR 1 . ^^^ This study demonstrates for the first time that VEGF B , in part through its receptor VEGF R 1 , promotes angiogenesis in association with an activation of Akt and eNOS related pathways . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The phosphorylation of Akt and its downstream effector endothelial NO synthase ( eNOS ) is pivotal to VEGF induced angiogenesis . ^^^ We therefore investigated the effect of Ang 2 on VEGF induced Akt and eNOS phosphorylation . ^^^ Ang 2 diminished the VEGF induced phosphorylation of Akt and eNOS in endothelial cells , whereas the autophosphorylation of VEGF receptors was unaffected . ^^^ Treatment of endothelial cells with pertussis toxin ( PTX ) totally abolished the AT 2 receptor mediated inhibition of VEGF induced endothelial cell migration and blocked the inhibition of Akt and eNOS phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Akt is a major activator of the endothelial nitric oxide synthase ( eNOS ) enzyme , but its potential role in intrahepatic resistance in cirrhosis is unknown . ^^^ Protein abundance and phosphorylation status of Akt and eNOS were examined by Western blotting . ^^^ RESULTS : The liver of cirrhotic animals showed a significant reduction in Akt and eNOS phosphorylation . ^^^ Adenoviral delivery of myr Akt restored eNOS phosphorylation and increased the intrahepatic concentration of guanosine 3 ' , 5 ' cyclic monophosphate . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Flow stimulated VEGFR 2 recruits phosphoinositide 3 kinase and mediates activation of Akt and eNOS . ^^^ Inhibiting VEGFR 2 kinase with selective inhibitors blocks flow induced activation of Akt and eNOS and production of NO . ^^^ Decreasing VEGFR 2 expression with antisense VEGFR 2 oligonucleotides significantly attenuates activation of Akt and eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
There have been conflicting results as to which protein kinases protein kinase A , protein kinase B ( Akt ) , other Ser / Thr protein kinases , or tyrosine kinases are responsible for shear dependent eNOS regulation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Further studies have revealed that HDL stimulates eNOS through src and PI 3 kinase mediated signaling , which leads to parallel activation of Akt and mitogen activated protein kinases and their resultant independent modulation of the enzyme . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Using phospho specific antibodies , we observed that either adiponectin or insulin treatment ( but not LPA treatment ) caused phosphorylation of both Akt at Ser 473 and endothelial nitric oxide synthase ( eNOS ) at Ser 1179 that was inhibitable by wortmannin . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Stretch induced phosphorylation of Akt and eNOS in lung endothelial cells was demonstrated by immunohistochemistry and increased NO production by in situ fluorescence microscopy . ^^^ These findings demonstrate that circumferential stretch activates NO production in pulmonary endothelial cells by a signaling cascade involving phosphatidylinositol 3 OH kinase , Akt , and eNOS and that this response is independent from the mechanical factors causing vascular distension . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
VEGF pretreatment of cultured endothelial cells also markedly potentiated S1P promoted eNOS phosphorylation at Ser 1179 , as well as S1P mediated activation of kinase Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Total and phosphorylated protein kinase B ( Akt ) as well as total and phosphorylated eNOS were quantitatively assessed in mice penes using Western immunoblot and immunohistochemistry . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Here , using a model in which we have utilized isolated primary endothelial cells , we demonstrate that ET 1 binding to sinusoidal endothelial cell ETB receptors led to increased protein kinase B / Akt phosphorylation , endothelial cell nitric oxide synthase ( eNOS ) phosphorylation , and NO synthesis . ^^^ Furthermore , eNOS activation was not dependent on tyrosine phosphorylation , and pretreatment of endothelial cells with pertussis toxin as well as overexpression of a dominant negative G protein coupled receptor kinase construct that sequesters betagamma subunits inhibited Akt phosphorylation and NO synthesis . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
CsA treatment rapidly led to an increase in myocardial Hsp 90 expression promoting the recruitment of Akt and calcineurin , thereby promoting eNOS activation through Ser 1177 phosphorylation and Thr 495 dephosphorylation , respectively . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This study examines the notion that heat shock protein ( HSP ) 90 binding to nitric oxide ( NO ) , endothelial NO synthase ( eNOS ) , and PI3K Akt regulate angiopoietin ( Ang ) 1 induced angiogenesis in porcine coronary artery endothelial cells ( PCAEC ) . ^^^ Ang 1 also led to stimulation of HSP 90 binding to eNOS and a significant increase in Akt phosphorylation . ^^^ We conclude that stimulated HSP 90 binding to eNOS and activation of the PI 3 Akt pathway contribute to Ang 1 induced eNOS phosphorylation , NO production , and angiogenesis in PCAEC . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Adiponectin promoted the phosphorylation of AMP activated protein kinase ( AMPK ) , protein kinase Akt / protein kinase B , and endothelial nitric oxide synthesis ( eNOS ) in HUVECs . ^^^ Transduction with either dominant negative AMPK or dominant negative Akt abolished adiponectin induced eNOS phosphorylation as well as adiponectin stimulated HUVEC migration and differentiation . ^^^ Dominant negative Akt or the phosphatidylinositol 3 kinase inhibitor LY 294002 blocked adiponectin stimulated Akt and eNOS phosphorylation , migration , and differentiation without altering AMPK phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Celiprolol activates eNOS through the PI3K Akt pathway and inhibits VCAM 1 Via NF kappaB induced by oxidative stress . ^^^ Because it was recently reported that phosphatidylinositol 3 kinase ( PI3K ) and its downstream effector Akt are implicated in the activation of eNOS and that regulation of VCAM 1 expression is mediated via nuclear factor kappaB ( NF kappaB ) , we hypothesized that celiprolol activates phosphorylation of eNOS through the PI3K Akt signaling pathway ; that celiprolol modulates VCAM 1 expression , which is associated with inhibiting NF kappaB phosphorylation ; and that celiprolol suppresses NAD ( P ) H oxidase p22phox , p47phox , gp91phox , and nox 1 expression in the left ventricle of deoxycorticosterone acetate ( DOCA ) salt hypertensive rats . eNOS and Akt phosphorylation upregulated by celiprolol alone were suppressed by treatment with celiprolol plus wortmannin . ^^^ In conclusion , celiprolol suppresses VCAM 1 expression because of inhibition of oxidative stress , NF kappaB , and signal transduction , while increasing eNOS via stimulation of the PI3K Akt signaling pathway and improving cardiovascular remodeling . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This was accompanied by the attenuation of troglitazone stimulated increases in the phosphorylation of Akt and eNOS Ser ( 1179 ) with no alteration in eNOS Ser ( 116 ) dephosphorylation . ^^^ We also found that bisphenol A diglycidyl ether , a PPARgamma antagonist , partially inhibited troglitazone stimulated NO production with a concomitant reduction in VEGF KDR / Flk 1 Akt mediated eNOS Ser ( 1179 ) phosphorylation but with no alteration in eNOS Ser ( 116 ) dephosphorylation induced by troglitazone . ^^^ Taken together , our results demonstrate that prolonged treatment with troglitazone increases endothelial NO production by at least two independent signaling pathways : PPARgamma dependent , VEGF KDR / Flk 1 Akt mediated eNOS Ser ( 1179 ) phosphorylation and PPARgamma independent , eNOS Ser ( 116 ) dephosphorylation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Insulin activated the protein kinase Akt in cultured endothelial cells and increased the phosphorylation of eNOS on Ser ( 1177 ) but failed to increase endothelial cyclic GMP levels or to elicit the relaxation of endothelium intact porcine coronary arteries . ^^^ In platelets , insulin also elicited the activation of Akt as well as the phosphorylation of eNOS and initiated NO production which was associated with increased cyclic GMP levels and the inhibition of thrombin induced aggregation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Increased cytosolic Ca2+ or activation of the kinase Akt leads to eNOS activation and its dissociation from caveolin 1 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Inhibition of VEGFR 2 blocked activation of extracellular regulated kinase , p 38 , Akt , and endothelial nitric oxide synthetase ( eNOS ) by VEGF , but did not inhibit p 38 activation by the VEGFR 1 specific ligand , placental growth factor ( PIGF ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Western blot analysis for phosphorylated ERK1 / 2 , Akt , p 38 mitogen activated protein kinase ( MAPK ) , and eNOS showed that shear stress rapidly increased phosphorylation of ERK1 / 2 and Akt but not of p 38 MAPK . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Treatment with EGCG induced a sustained activation of Akt , ERK1 / 2 , and eNOS Ser 1179 phosphorylation . ^^^ Simultaneous treatment of cells with selective inhibitors for cAMP dependent protein kinase ( PKA ) and Akt completely prevented the increase in eNOS activity by EGCG after 15 min , indicating that both kinases act in concert . ^^^ Akt inhibition prevented eNOS Ser 1179 phosphorylation , whereas inhibition of PKA did not influence Akt and eNOS Ser 1179 phosphorylation . ^^^ Pretreatment of endothelial cells with EGCG for 4 h markedly enhanced the increase in eNOS activity stimulated by Ca ionomycin , suggesting that Akt accounts for prolonged eNOS activation . ^^^ Our results indicate that EGCG induced endothelium dependent vasodilation is primarily based on rapid activation of eNOS by a phosphatidylinositol 3 kinase , PKA , and Akt dependent increase in eNOS activity , independently of an altered eNOS protein content . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Hypoxia increases Hsp 90 binding to eNOS via PI3K Akt in porcine coronary artery endothelium . ^^^ Hypoxia also led to a significant increase in Akt phosphorylation and upregulation of Hsp 90 binding to eNOS . ^^^ Pretreatment of cells with either 1 microg / ml geldanamycin ( a specific inhibitor of Hsp 90 ) or 500 nM wortmannin ( a specific PI 3 kinase inhibitor ) suppressed hypoxia stimulated Akt and eNOS phosphorylation and significantly attenuated hypoxia stimulated Hsp 90 binding to eNOS . ^^^ These data demonstrate that hypoxia leads to increased eNOS phosphorylation via stimulated Hsp 90 binding to eNOS and activation of the PI 3 Akt pathway . ^^^ We conclude that a coordinated interaction between Hsp 90 and PI 3 Akt may be an important mechanism by which eNOS activity and NO production is upregulated in hypoxic heart . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The phosphorylation of downstream molecules of Akt , forkhead transcription factors ( FKHR ) , endothelial nitric oxide synthase ( eNOS ) , and glycogen synthase kinase 3beta ( GSK 3beta ) was also increased . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Compared with HUVEC WT , in HUVEC G972R after 2 and 10 minutes of insulin stimulation , IRS 1 associated PI 3 K activity was reduced by 47 % and 32 % , respectively ; Akt phosphorylation was decreased by 40 % at both time points ; and eNOS Ser 1177 phosphorylation was reduced by 38 % and 51 % , respectively . ^^^ CONCLUSIONS : Our data demonstrate that genetic impairment of the ( IRS ) 1 / PI3 K / PDK 1 / Akt insulin signaling cascade determines impaired insulin stimulated NO release and suggest that the G972R IRS 1 polymorphism , through a direct impairment of Akt / eNOS activation in endothelial cells , may contribute to the genetic predisposition to develop endothelial dysfunction and cardiovascular disease . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The eNOS induced mechanical effects were paralleled with elevated phosphorylation of Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Here we show that HDL stimulates NO release in human endothelial cells and induces vasodilation in isolated aortae via intracellular Ca2+ mobilization and Akt mediated eNOS phosphorylation . ^^^ All three elevated intracellular Ca2+ concentration and activated Akt and eNOS , which resulted in NO release and vasodilation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Recent research suggests that endothelial TF expression is positively regulated by RhoA and p38mapk , but negatively by Akt / endothelial nitric oxide synthase ( eNOS ) pathway . ^^^ Activation of RhoA but not p38mapk by thrombin was prevented by rHDL . rHDL stimulated Akt / eNOS pathway . ^^^ The phosphatidylinositol 3 kinase ( PI3K ) inhibitors wortmannin or LY 294002 abolished the activation of Akt / eNOS and reversed the inhibitory effect of rHDL on TF expression . ^^^ In conclusion , rHDL inhibits thrombin induced human endothelial TF expression through inhibition of RhoA and activation of PI3K but not Akt / eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Insulin stimulates NO production by a pathway involving IRS 1 / PI3 kinase / Akt / endothelial NO synthase ( eNOS ) . ^^^ This event was associated with impaired IRS 1 phosphorylation at Tyr 612 and Tyr 632 , two sites essential for engaging the p 85 subunit of PI 3 kinase , resulting in defective activation of PI 3 kinase , Akt , and eNOS . ^^^ Our data suggest that AII , acting via the type 1 receptor , increases IRS 1 phosphorylation at Ser 312 and Ser 616 via JNK and ERK1 / 2 , respectively , thus impairing the vasodilator effects of insulin mediated by the IRS 1 / PI 3 kinase / Akt / eNOS pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
BACKGROUND : Insulin increases endothelial nitric oxide ( NO ) production by activating endothelial nitric oxide synthase ( eNOS ) through protein kinase B ( Akt ) mediated phosphorylation of serine residue 1179 ( p eNOS serine 1179 ) . ^^^ Because fatty acids modulate insulin stimulated Akt signaling cascades in smooth muscle cells , we hypothesized that fatty acids would differentially regulate endothelial Akt signaling , eNOS phosphorylation , and NO production . ^^^ Activation and phosphorylation of Akt and eNOS were determined by immunoblotting . ^^^ RESULTS : Insulin stimulated Akt phosphorylation , eNOS phosphorylation , and NO production . ^^^ Treatment with the omega 3 fatty acid 20 : 5 , but not 18 : 1 , enhanced insulin stimulated NO production but failed to alter insulin stimulated Akt activation or eNOS serine 1179 phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Statin stimulation of BAECs increased eNOS phosphorylation at Ser 1179 and Ser 617 , which was blocked by the phosphatidylinositol 3 kinase ( PI 3 kinase ) / Akt inhibitor wortmannin , and at Ser 635 , which was blocked by the protein kinase A ( PKA ) inhibitor KT 5720 . ^^^ In conclusion , these data demonstrate that eNOS is acutely activated by statins independent of HMG CoA reductase inhibition and that in addition to Ser 1179 , eNOS phosphorylation at Ser 635 and Ser 617 through PKA and Akt , respectively , may explain , in part , a mechanism by which eNOS is activated in response to acute statin treatment . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
An increased Akt activity preceded the postischemic eNOS up regulation . ^^^ Intracerebroventricular injection ( i . c . v . ) of wortmannin , an inhibitor of phosphatidylinositol 3 kinase ( PI 3K ) , significantly inhibited the increases in both eNOS mRNA and its protein with concomitant inhibition of Akt activation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Phosphorylation of endothelial NO synthase ( eNOS ) increases NO production and reduces apoptosis through the Akt signaling pathway . ^^^ To elucidate the Akt signaling pathway involved in the opening and antiapoptotic effect of mitoKATP channel during delayed pharmacological preconditioning , the mitoKATP channel opener diazoxide ( DE , 7 microg / kg i . p . ) alone or DE plus Nomega nitro L arginine methyl ester ( L NAME , 30 microg / kg i . v . ) , an inhibitor of NOS , or wortmannin ( WTN , 15 microg / kg i . v . ) , an inhibitor of phosphatidylinositol 3 ' kinase ( PI 3 kinase ) , was administered to wild type ( WT ) or eNOS ( / ) mice during DE treatment . ^^^ Treatment with DE resulted in a 2 . 2 fold increase in phosphorylation of Akt and a significant increase in eNOS and inducible NOS ( iNOS ) proteins . ^^^ It is concluded that DE activates Akt through the PI 3 kinase signaling pathway and iNOS and eNOS is downstream of Akt . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
SDF 1alpha gene transfer did not affect ischemia induced expression of vascular endothelial growth factor ( VEGF ) but did enhance Akt and endothelial nitric oxide synthase ( eNOS ) activity . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Furthermore , we found that endothelial nitric oxide synthase ( eNOS ) , one of the Akt substrates , was highly expressed in LNCaP but not in other cells . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We further documented in vivo and in cultured endothelial cells that the cardioprotective effects of Hsp 90 were associated to its capacity to act as an adaptor for both the kinase Akt and the phosphatase calcineurin , thereby promoting eNOS serine 1177 phosphorylation and threonine 495 dephosphorylation , respectively . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Chronic hypoxia impaired eNOS phosphorylation and Akt phosphorylation under both the nonstimulated and substance P stimulated conditions , and 3 microM DEX restored these phosphorylations . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Regulation of erectile function may not be mediated exclusively by neurally derived NO : Blood flow induced fluid shear stress in the penile vasculature stimulates phosphatidyl inositol 3 kinase to phosphorylate protein kinase B , which in turn phosphorylates eNOS to generate NO . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Vascular smooth muscle cell ( VSMC ) proliferation , apoptotic cell death , together with Akt induction , telomerase activity , p27kip1 , and endothelial nitric oxide synthase ( eNOS ) expression was assessed in isolated arteries . ^^^ The molecular mechanisms underlying these responses seem to be a reduced Akt and eNOS activity . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Isolated perfused rat hearts were subjected to : ( a ) 35 minutes of ischemia and 120 minutes of reperfusion , and infarct size was determined by tetrazolium staining ; or ( b ) 35 minutes of ischemia and 7 minutes of reperfusion , and the phosphorylation states of Akt , endothelial NO synthase ( eNOS ) , and p70S6K were determined . ^^^ Western blot analysis demonstrated that Postcond induced a significant increase in phosphorylation of Akt , eNOS , and p70S6K in an LY and Wort sensitive manner . ^^^ In conclusion , we show for the first time that ischemic Postcond protects the myocardium by activating the prosurvival kinases PI3K Akt , eNOS , and p70S6K in accordance with the RISK pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
IBOP costimulation suppressed nitric oxide ( NO ) release from VEGF treated EC through decreased activation of Akt , eNOS , and PDK 1 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Carbon monoxide protects against cardiac ischemia reperfusion injury in vivo via MAPK and Akt eNOS pathways . ^^^ Exposure to 1000 ppm CO resulted in the activation of p 38 mitogen activated protein kinase ( p38MAPK ) , protein kinase Balpha ( Akt ) , endothelial nitric oxide synthase ( eNOS ) , and cyclic guanosine monophosphate ( cGMP ) in the myocardium . ^^^ CONCLUSIONS : CO has beneficial effects on cardiac ischemia reperfusion injury ; this effect is mediated by p38MAPK pathway and Akt eNOS pathway , including production of cGMP . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Black tea polyphenol induced eNOS activation appeared dependent upon the phosphatidylinositol 3 kinase Akt pathway , as it was significantly inhibited by LY 294002 and a dominant negative Akt , respectively . ^^^ Finally , a constitutively active mutant of MKK6bE , an upstream kinase for p 38 MAPK , enhanced both the basal and stimulated activity of Akt , leading to increased eNOS activity . ^^^ Taken together , these data identify the p 38 MAPK as an upstream component of Akt mediated eNOS activation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The former effect is mediated by pertussis toxin sensitive receptors ( possibly S1P ( 1 ) ) located on the endothelium and stimulating phosphatidylinositol 3 kinase / Akt / endothelial nitric oxide synthase ( eNOS ) . ^^^ Finally , S1P also protects endothelial cells from apoptosis through activation of phosphatidylinositol 3 kinase / Akt / eNOS via S1P ( 1 ) and S1P ( 3 ) receptors . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The levels of Akt , Bad , and eNOS ( total and phosphorylated ) proteins were measured . ^^^ Phosphorylation pattern of eNOS supported the involvement of PI3K / Akt pathway in this process . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In the present study , we show that Cy3G regulates phosphorylation of eNOS and Akt , affects the interaction between eNOS and soluble guanylyl cyclase , and increases cyclic guanosine monophosphate ( cGMP ) production . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Ad . hTK upregulated eNOS mRNA and protein and activated Akt B through Ser 473 phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The vascular endothelial growth factor ( VEGF ) contributes to activation of eNOS by Ca2+ / calmodulin and also stimulates the protein kinase Akt , which directly phosphorylates eNOS on Ser 1177 and increases enzyme activity . ^^^ Western blot analysis and ELISAs were used to study the temporal profiles of Akt , phospho Akt at Ser 437 , eNOS , phospho eNOS at Ser 1177 , and VEGF expression , respectively . ^^^ Immunohistochemical studies were performed to examine the spatial expression patterns of phospho Akt at Ser 437 and phospho eNOS at Ser 1177 . ^^^ RESULTS : Increase in phospho Akt at Ser 437 was observed transiently 0 . 5 to 2 hours after reperfusion , whereas elevation of phospho eNOS at Ser 1177 and VEGF expression was observed from 6 hours after reperfusion . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Both estradiol and LiCl enhanced the expression of eNOS mRNA with the phosphorylation of GSK 3beta , but not Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The effect of CsA was not related to inhibition of two known eNOS kinases , protein kinase B ( Akt ) and protein kinase A , because CsA did not affect Akt or protein kinase A activation . ^^^ In rabbit aorta perfused ex vivo , CsA also significantly inhibited flow induced eNOS phosphorylation at Ser 1179 but had no effect on Akt measured by phosphorylation at Ser 473 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
MPA attenuated the E 2 induced phosphorylation of Akt , a kinase that phosphorylates eNOS . ^^^ Treatment with pure progesterone receptor ( PR ) antagonist RU 486 completely abolished the inhibitory effect of MPA on E 2 induced Akt phosphorylation and eNOS phosphorylation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We examined the hypothesis that this is due to activation of Akt / PKB which subsequently increases eNOS activity . ^^^ METHODS AND RESULTS : Treatment of bovine microvascular and human umbilical endothelial cells ( HUVEC ) with 17 beta estradiol ( E 2 ) ( 10 ( 9 ) to 10 ( 5 ) M ) increased phosphorylation of Akt within 1 min and this was followed by phosphorylation of eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The process requires Src and erk signaling and eNOS phosphorylation by phosphoinositide 3 kinase ( PI 3 kinase ) Akt kinase , with Src and PI 3 kinase associating with ERalpha upon ligand activation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
METHODS AND RESULTS : Because protein kinase Akt and endothelial nitric oxide synthase ( eNOS ) are critical for angiogenesis , we studied the effects of 17 AAG on the phosphorylation and expression of Akt and eNOS in human umbilical vein endothelial cells . ^^^ In a concentration and time dependent manner , inhibition of Hsp 90 by 17 AAG decreased Akt and eNOS expression by 74 % and 81 % , respectively . ^^^ Furthermore , treatment with 17 AAG decreased basal and vascular endothelial growth factor stimulated Akt and eNOS phosphorylation . ^^^ CONCLUSIONS : These findings indicate that Hsp 90 is important not only for Akt and eNOS phosphorylation but also for eNOS gene transcription and suggests that Hsp 90 may be a novel target for anti angiogenic therapy . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In parallel with NO production , 17beta estradiol treatment rapidly increased phosphorylation of both eNOS ( p eNOS ) and Akt ( p Akt ) . ^^^ PI 3 kinase inhibitors also blocked the latter effects ; together , these data are consistent with ER activation of the PI 3 kinase p Akt p eNOS pathway . ^^^ Long term estrogen exposure increased levels of cerebrovascular p Akt and p eNOS as well as basal NO production . ^^^ Thus , in addition to the rapid activation of PI 3 kinase , p Akt , and p eNOS , estrogen signaling via nontranscriptional , kinase mechanisms has long term consequences for vascular function . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The phosphatidylinositol 3 kinase ( PI 3 K ) pathway , which activates serine / threonine protein kinase Akt , enhances endothelial nitric oxide synthase ( eNOS ) phosphorylation and nitric oxide ( NO ) production . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
High glucose can trigger endothelial cell apoptosis by de activation of endothelial nitric oxide synthase ( eNOS ) . eNOS was recently demonstrated to be extensively regulated by Akt and heat shock protein 90 ( HSP 90 ) . ^^^ The protein interaction of eNOS / HSP90 and eNOS / Akt were studied in cultured human umbilical vein endothelial cells ( HUVECs ) exposed to either control level ( 5 . 5 mM ) or high level ( 33 mM ) glucose for different durations ( 2 , 4 , 6 , and 24 h ) . ^^^ The results showed that the protein interactions between eNOS and HSP 90 and between eNOS and Akt and the phosphorylation of eNOS were up regulated by high glucose exposure for 2 4 h . ^^^ During early hours of exposure , the protein interactions of eNOS / HSP90 and eNOS / Akt and the phosphorylation of eNOS were all inhibited by geldanamycin , an HSP 90 inhibitor . ^^^ LY 294002 , a phosphatidylinositol 3 ( PI 3 ) kinase inhibitor , inhibited the association of eNOS / Akt and the phosphorylation of eNOS but had no effect on the interaction between eNOS and HSP 90 during early hours of exposure . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Functional proteomic analysis of a three tier PKCepsilon Akt eNOS signaling module in cardiac protection . ^^^ Among 93 different PKCepsilon associated proteins that have been identified , Akt and endothelial nitric oxide ( NO ) synthase ( eNOS ) are of importance because of their independent abilities to promote cell survival and prevent cell death . ^^^ The simultaneous association of PKCepsilon , Akt , and eNOS has not been examined , and , in particular , the formation of a module containing these three proteins and the role of such a module in the regulation of NO production and cardiac protection are unknown . ^^^ Using recombinant proteins in vitro and PKCepsilon transgenic mouse hearts , we demonstrate the following : 1 ) PKCepsilon , Akt , and eNOS interact and form signaling modules in vitro and in the mouse heart . ^^^ Activation of either PKCepsilon or Akt enhances the formation of PKCepsilon Akt eNOS signaling modules . 2 ) PKCepsilon directly phosphorylates and enhances activation of Akt in vitro , and PKCepsilon activation increases phosphorylation and activation of Akt in PKCepsilon transgenic mouse hearts . 3 ) PKCepsilon directly phosphorylates eNOS in vitro , and this phosphorylation enhances eNOS activity . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
VEGF binding initiates tyrosine phosphorylation of KDR , which results in activation of downstream signaling enzymes including ERK1 / 2 , Akt and eNOS , which contribute to angiogenic related responses in EC . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Expression of endothelial nitric oxide synthase ( eNOS ) and Akt phosphorylation in response to AM ( 10 ( 7 ) mol / l ) were also diminished in OZ rats . ^^^ Fluvastatin restored the eNOS expression and Akt phosphorylation [ eNOS expression ( relative intensity ) : LZ , 2 . 3 + / 0 . 4 ; OZ , 1 . 0 + / 0 . 2 ; OZ + Flu , 1 . 8 + / 0 . 3 ; Akt phosphorylation ( relative intensity ) : LZ , 2 . 3 + / 0 . 2 ; OZ , 1 . 0 + / 0 . 3 ; OZ + Flu , 1 . 9 + / 0 . 2 ] . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This study was designed to test the hypothesis that ( a ) the cervix of pregnant rats expresses vascular endothelial growth factor ( VEGF ) and components of the angiogenic signaling pathway [ VEGF receptors ( Flt 1 , KDR ) , activity of protein kinase B , Akt ( phosphorylated Akt ) , and endothelial nitric oxide synthase ( eNOS ) ] and von Willebrand Factor ( vWF ) and that these molecules undergo changes with pregnancy , and ( b ) bilateral pelvic neurectomy ( BLPN ) alters levels of VEGF concentration in the cervix . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In vascular endothelial cells , the SHR for estrogen , estrogen receptor ( ER ) alpha , is targeted by unknown mechanisms to a functional signaling module in membrane caveolae that enables estrogen to rapidly activate the mitogen activated protein kinase and phosphatidylinositol 3 Akt kinase pathways , and endothelial NO synthase ( eNOS ) . ^^^ Disruption of complex formation between ERalpha and striatin blocks estrogen induced rapid activation mitogen activated protein kinase , Akt kinase , and eNOS , but has no effect on ER dependent regulation of an estrogen response element driven reporter plasmid . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Based on these observations , the present study utilizes laser Doppler flowmetry , histochemistry , enzyme immunoassay , immunoblotting , and real time PCR to characterize and compare the patterns of regional cerebral blood flow and synthesis of angiogenic molecules [ basic fibroblast growth factor ; nitric oxide synthase isoforms ( endothelial , neuronal and inducible ) ; vascular endothelial growth factor ( VEGF ) and its signaling molecules ( VEGF receptors , phosphorylated Akt , endothelial nitric oxide synthase eNOS ] between male SHRSP and SHR . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Akt phosphorylates endothelial NO synthase ( eNOS ) and thereby enhances endothelial NO synthesis and influences postnatal vessel growth . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Thus , atRA decreases eNOS Ser ( 1179 ) phosphorylation through a mechanism that depends on VEGF KDR / Flk 1 mediated Akt phosphorylation but is independent of RARE , leading to reduction in NO production . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
RWPs caused the sustained phosphorylation of Akt and eNOS at Ser 1177 in endothelial cells , which were abolished by MnTMPyP and inhibitors of PI 3 kinase . ^^^ These data demonstrate that RWPs induce the redox sensitive activation of the PI 3 kinase / Akt pathway in endothelial cells which , in turn , causes phosphorylation of eNOS , resulting in an increased formation of NO . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We showed previously that flow activated the phosphatidylinositol 3 kinase ( PI3K ) , Akt , and endothelial nitric oxide synthase ( eNOS ) via Src kinase dependent transactivation of vascular endothelial growth factor receptor 2 ( VEGFR 2 ) . ^^^ We found here that laminar flow ( shear stress = 12 dynes / cm2 ) rapidly stimulated Gab 1 tyrosine phosphorylation in both bovine aortic endothelial cells and human umbilical vein endothelial cells , which correlated with activation of Akt and eNOS . ^^^ Gab 1 phosphorylation as well as activation of Akt and eNOS by flow was inhibited by the Src kinase inhibitor PP 2 ( 4 amino 5 ( 4 chlorophenyl ) 7 ( t butyl ) pyrazolo [ 3 , 4 d ] pyrimidine ) and VEGFR 2 kinase inhibitors SU 1498 and VTI , suggesting that flow mediated Gab 1 phosphorylation is Src kinase dependent and VEGFR 2 dependent . ^^^ Furthermore , transfection of a Gab 1 mutant lacking p 85 binding sites inhibited flow induced activation of Akt and eNOS . ^^^ Finally , knockdown of endogenous Gab 1 by small interference RNA abrogated flow activation of Akt and eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
CONCLUSION : Adenosine protects mitochondria from oxidant damage through a pathway involving A ( 2 ) receptors , eNOS , NO , PI 3 kinase / Akt , and Src tyrosine kinase . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Sections from the implants were stained with antibodies against vascular endothelial growth factor receptor 2 ( VEGFR 2 ) , Akt , phosphorylated Akt ( p Akt ) , endothelial nitric oxide synthase ( eNOS ) , phosphorylated eNOS ( p eNOS ) , inducible NOS ( iNOS ) , and 3 nitrotyrosine ( 3 NT , a marker for nitrosylated proteins ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We found that treatment of EC with a low dose of pitavastatin induced eNOS phosphorylation at Ser 1177 , activated Akt phosphorylation at Ser 473 in a time and dose dependent manner , and increased NO production . ^^^ These results suggest that the activation of eNOS with a low dose of pitavastatin ( 0 . 1 microM ) involves phosphoinositide 3 kinase and the Akt pathway and produces NO in EC , which is dependent on post transcriptional regulation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Besides acting as an allosteric enhancer , Hsp 90 was shown to serve as a module recruiting Akt to phosphorylate the serine 1179 / 1177 ( bovine / human ) residue of eNOS . ^^^ To address this issue , we treated bovine eNOS stably transfected human embryonic kidney 293 cells with Hsp 90 inhibitors and determined the alterations of phospho eNOS , Akt , and PDK 1 . ^^^ In Hsp 90 inhibited cells , eNOS associated phospho Akt was decreased , but the total amount of Akt associated with eNOS remained the same . ^^^ Silencing the PDK 1 gene by small interfering RNA was sufficient to induce reduction of phospho Akt and consequent loss of serine 1179 phosphorylated eNOS . ^^^ Moreover , overexpression of PDK 1 , but not Akt , reversed Hsp 90 inhibition induced loss of eNOS serine 1179 phosphorylation and salvaged enzymatic activity . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Expression of NADPH oxidase , Akt , pAkt , Bcl 2 , Bax , IkappaB , caveolin 1 and eNOS was evaluated by Western blot analysis . 2 ET 1 significantly enhanced ROS generation and cell proliferation following 24 h incubation , both of which were prevented by BQ 788 or apocynin , consistent with the ability of ET 1 to directly upregulate NADPH oxidase . ^^^ Furthermore , ET 1 downregulated expression of caveolin 1 and eNOS , which was attenuated by BQ 788 or apocynin . 3 In summary , our results suggest that ET 1 affects oxidative stress , proliferation and apoptosis possibly through ET ( B ) , NADPH oxidase , Akt , Bax and caveolin 1 mediated mechanisms . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Up regulation of Akt and eNOS induces vascular smooth muscle cell differentiation in hypertension in vivo . ^^^ Both drugs also significantly reduced aortic NAD ( P ) H oxidase activity and p38MAPK and ERK expression , whereas p Akt , eNOS , and SM 2 were significantly increased in SHRSP aortas . ^^^ Furthermore , E 4177 was more effective than cilazapril at inducing VSMC differentiation by reducing NAD ( P ) H oxidase activity , and up regulating p Akt , eNOS , and SM 2 . ^^^ Thus , an ACE inhibitor and an AT 1 receptor antagonist inhibited VSMC dedifferentiation through inhibition of NAD ( P ) H oxidase activity and up regulation of eNOS and Akt in SHRSP aortas , suggesting that in contrast to the in vitro experiments , AT 1 receptor mediated NAD ( P ) H oxidase generated ROS , eNOS , and Akt might be crucial determinants for the VSMC phenotype in hypertension in vivo . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Activation of Akt and eNOS by HF was completely blocked by the phosphatidylinositol 3 kinase ( PI 3 kinase ) inhibitor , LY 294002 ( 10 micromol / L ) . ^^^ CONCLUSIONS : Inhibition of Rho kinase leads to the activation of the PI 3 kinase / Akt / eNOS pathway and cardiovascular protection . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Recent reports show that H ( 2 ) O ( 2 ) induces phosphorylation and activation of endothelial nitric oxide synthase ( eNOS ) through an Akt phosphorylation dependent pathway . ^^^ In this study , we assessed activation of eNOS and Akt by determining their phosphorylation status . ^^^ Whereas moderate levels of H ( 2 ) O ( 2 ) ( 100 microM ) activated the Akt / eNOS pathway , higher levels ( 500 microM ) did not , suggesting differential effects by differing levels of oxidative stress . ^^^ However , down regulation of cell membrane surface and intracellular free thiols was associated with the inhibition of phosphorylation , suggesting that oxidation of non GSH thiols inhibits the H ( 2 ) O ( 2 ) induced phosphorylation of eNOS and Akt . ^^^ Similarly , DEM and CDNB inhibited TNF alpha induced Akt and eNOS phosphorylation , suggesting that thiol modification is involved in eNOS inductive pathways . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Taken together , these results indicate that HGF not only phosphorylates eNOS through the PI3K / Akt pathway , but also partially through the MAPK pathway , and that these two pathways may interact . ^^^ Our study thus demonstrates a novel activity of HGF the stimulation of NO production which occurs via eNOS phosphorylation that may in turn be mediated by cross talk between the PI3K / Akt and MAPK pathways . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The aortic levels of total eNOS , phosphorylated eNOS at Ser 1177 ( p eNOS ) , total Akt , and phosphorylated Akt at Ser 473 ( p Akt ) were increased in 2K1C mice on day 14 , whereas only eNOS levels were increased on day 42 . ^^^ Administration of nicardipine for 4 days before the excision of aortas 14 days after clipping not only reduced blood pressure but also decreased the aortic levels of eNOS , p eNOS , Akt , p Akt , and cGMP to sham levels , whereas the administration of PD 123319 or icatibant to 2K1C mice decreased p eNOS and cGMP to sham levels without affecting blood pressure and the levels of eNOS , Akt and p Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In aged , but not young rats , sildenafil prolonged erection and increased the protein expressions of phosphorylated endothelial NO synthase ( eNOS ) at serine 1177 and phosphorylated Akt at serine 473 in penes . ^^^ We conclude that erectile ability can be enhanced under preconditions of erectile impairment by long term inhibition of phosphodiesterase 5 and that the effect is mediated by Akt dependent eNOS phosphorylation . ^^^ The lack of erectile ability enhancement in young rats by long term phosphodiesterase 5 inhibition may relate to restrained NO signaling by phosphodiesterase 5 up regulation , lack of incremental Akt and eNOS phosphorylation , and heightened Rho kinase signaling in the penis . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
METHODS AND RESULTS : We demonstrate that insulin mediated tyrosine phosphorylation of IRS 1 and serine phosphorylation of Akt , eNOS , and NO production are significantly inhibited by treatment of bovine aortic endothelial cells with 100 micromol / L FFA composed of palmitic acid for 3 hours before stimulation with 100 nM insulin . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Conversely , constitutively active MKK 6 induced p 38 MAPK activation that recapitulated the effects of polyphenols by inducing ERalpha phosphorylation and downstream activation of Akt , and eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Stimulated HSP 90 binding to eNOS and activation of the PI 3 Akt pathway contribute to globular adiponectin induced NO production : vasorelaxation in response to globular adiponectin . ^^^ The present study examined potential interactions between endothelial NO synthase ( eNOS ) , heat shock protein ( HSP ) 90 , and Akt in vascular endothelial cells stimulated with globular adiponectin to produce nitric oxide ( NO ) . ^^^ Globular adiponectin induced eNOS phosphorylation was accompanied by eNOS HSP 90 Akt complex formation , resulting in a dose dependent increase in NO release . ^^^ These results indicate that stimulated HSP 90 binding to eNOS and activation of the PI 3 Akt pathway contribute to globular adiponectin induced eNOS phosphorylation and NO production , and to endothelium dependent vasorelaxation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The increased expression and phosphorylation of eNOS with RPM appears to be regulated by mechanisms other than Akt or Caveolin 1 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Insulin stimulated protein kinase B ( akt ) and endothelial nitric oxide synthase ( eNOS ) phosphorylation were preserved in arteries from diabetic mice ; however , eNOS protein dimers were markedly diminished . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
High glucose induced apoptosis in human vascular endothelial cells is mediated through NF kappaB and c Jun NH 2 terminal kinase pathway and prevented by PI3K / Akt / eNOS pathway . ^^^ In this study we further elucidated the roles of the transcriptional factor NF kappaB , phosphatidylinositol 3 ' kinase ( PI3K ) , Akt and endothelial nitric oxide synthase ( eNOS ) in the apoptosis of HUVECs induced by high glucose . ^^^ Activation of the ROS / PI3K / Akt / eNOS signaling pathway in early phase exerts protective effects against the induction of apoptosis by high glucose . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Further mechanistic studies demonstrated that the effects of myristic acid on eNOS function were not dependent on PI 3 kinase , Akt kinase , or calcium . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Here , we show that exposure of bovine aortic endothelial cells to high glucose ( 25 mM ) for 24 h impaired insulin mediated tyrosine phosphorylation of IRS 1 , serine phosphorylation of Akt , activation of eNOS , and production of NO . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
METHODS : Proinflammatory cytokine levels , Akt and NOS activities , eNOS phosphorylation , and NOS expressions were assessed in aorta from norfloxacin treated and untreated cirrhotic rats . ^^^ RESULTS : Aortic eNOS and iNOS protein expressions , Akt activity , and eNOS phosphorylation by Akt at serine 1177 were up regulated in cirrhotic rats . ^^^ Norfloxacin administration significantly decreased the incidence of gram negative translocation and proinflammatory cytokine ( tumor necrosis factor alpha , interferon gamma , and interleukin 6 ) levels ; norfloxacin also decreased aortic Akt activity , eNOS phosphorylation , and NOS expressions and activities . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Increasing shear stress from 3 to 15 dynes / cm2 very rapidly increased eNOS Ser 1177 , ERK1 / 2 ( extracellular signal regulated kinase 1 and 2 ) and Akt , but not p 38 MAPK ( p 38 mitogen activated protein kinase ) phosphorylation by Western analysis . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The vascular diameter was correlated with phosphorylation of Akt and serine 1177 residue of eNOS , and formation of NO bound guanylate cyclase ( GC ) by immuoflorescence study . ^^^ The constriction during reperfusion after 45 min of ischemia is supposedly caused by the inhibition of Akt mediated eNOS Ser 1177 phosphorylation , which was suppressed by a PKC inhibitor chelerythrine , or ROS scavengers N 2 mercaptopropionyl glycine ( MPG ) and 4 , 5 Dihydroxy 1 , 3 benzenedisulfonic acid disodium salt ( Tiron ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
ATRA as well as Am 580 enhanced endothelial NO synthase ( eNOS ) phosphorylation at Ser 1177 as well as Akt phosphorylation at Ser 473 without changing their protein expression . ^^^ Overexpression of dominant negative Akt inhibited the eNOS phosphorylation . ^^^ CONCLUSIONS : ATRA increases NO production by eNOS phosphorylation through RAR mediated PI3K / Akt pathway activation in vascular ECs and possibly plays beneficial roles in vascular endothelium . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Phosphorylation of Akt , a mediator of shear stress induced eNOS phosphorylation at Ser 1177 , was decreased in the diabetic penis at baseline , but it was restored by ES . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
However , the changes of Akt phosphorylation and eNOS activity in endothelial cells in response to lipoprotein treatments under our assay condition were not significant . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Glimepiride activates eNOS with a mechanism Akt but not caveolin 1 dependent . ^^^ Caveolin 1 silencing did not change eNOS and Akt phosphorylation induced by glimepiride . ^^^ These findings suggest that glimepiride induces eNOS phosphorylation in endothelial cells through an Akt dependent mechanism , not regulated by caveolin 1 . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Role of PECAM 1 in the shear stress induced activation of Akt and the endothelial nitric oxide synthase ( eNOS ) in endothelial cells . ^^^ Endothelial cell stimulation with shear stress elicited the phosphorylation of Akt and eNOS as well as of the AMP activated protein kinase ( AMPK ) . ^^^ While the shear stress induced tyrosine phosphorylation of PECAM 1 as well as the serine phosphorylation of Akt and eNOS were abolished by the pre treatment of cells with the tyrosine kinase inhibitor PP 1 the phosphorylation of AMPK was unaffected . ^^^ Down regulation of PECAM 1 using a siRNA approach attenuated the shear stress induced phosphorylation of Akt and eNOS , as well as the shear stress induced accumulation of cyclic GMP levels while the shear stress induced phosphorylation of AMPK remained intact . ^^^ A comparable attenuation of Akt and eNOS ( but not AMPK ) phosphorylation and NO production was also observed in endothelial cells generated from PECAM 1 deficient mice . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Akt and endothelial nitric oxide synthase ( eNOS ) protein expression and their phosphorylated forms were also evaluated by Western blot . ^^^ As a consequence , Akt phosphorylation , eNOS protein and its phosphorylated form were significantly higher in the samples from non diabetic patients compared with diabetic patients . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Phosphorylation of the serine threonine kinase Akt , which activates eNOS , was substantially reduced in sinusoidal endothelial cells from injured livers . ^^^ Thus , an important mechanism underlying impaired activity of eNOS in injured sinusoidal endothelial cells is defective phosphorylation of Akt caused by overexpression of GRK 2 after injury . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
VEGF induced eNOS phosphorylation was abolished by SU 1498 , a VEGF R 2 inhibitor , LY 294002 , a PI3K inhibitor , and infection of cells with an adenovirus carrying a dominant negative mutant of Akt , demonstrating the requirement of the VEGF R2 / IRS 1 / PI3K / Akt axis for activation of eNOS . ^^^ CONCLUSIONS : Our data demonstrate that in GENC , VEGF stimulates VEGF R2 / IRS 1 / PI3K / Akt axis to regulate eNOS phosphorylation on Ser 1177 in conjunction with the ERK signaling pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Because of the importance of endothelial nitric oxide synthase ( eNOS ) in vascular and platelet function , we hypothesized that losartan and its metabolites would stimulate eNOS and its upstream activators Akt and phosphatidylinositol 3 kinase ( PI3K ) . ^^^ Treatment of endothelial cells ( ECs ) with losartan and both metabolites stimulated phosphorylation of Akt and eNOS in the absence of angiotensin 2 . ^^^ In endothelium of intact rat aorta , EXP 3179 also stimulated Akt and eNOS phosphorylation . ^^^ CONCLUSIONS : The losartan metabolite EXP 3179 stimulates eNOS phosphorylation and suppresses tumor necrosis factor alpha induced EC apoptosis by activating the VEGFR2 / PI3K / Akt pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Western analysis demonstrated that FAKab prevented flow induced phosphorylation of FAK ( pY 397 FAK ) , Akt ( pS 473 Akt ) , and eNOS ( pS 1179 eNOS ) . ^^^ Inhibition of FAK signaling with FAKab impaired FID and phosphorylation of Akt and eNOS . ^^^ Our data suggest that the activation of FAK is central to the mechanotransduction of FID via regulation of activation of Akt and eNOS . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Expression of eNOS , HSP 90 , caveolin 1 , Akt , phosphorylated eNOS ( eNOS Ser 1177 ) , and GTPCH 1 were determined by Western blot analysis . ^^^ Expression of eNOS , caveolin 1 , phosphorylated Akt , and eNOS Ser 1177 was enhanced in aortas exposed to increased blood flow . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Although impaired vascular maturation in Akt 1 ( / ) mice may be attributed to reduced activation of endothelial nitric oxide synthase ( eNOS ) , the major phenotypic changes in vascular permeability and angiogenesis were linked to reduced expression of two endogenous vascular regulators , thrombospondins 1 ( TSP 1 ) and 2 ( TSP 2 ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In endothelial cells , S1P has been shown to modulate the activity of the endothelial nitric oxide synthase ( eNOS ) through phosphorylation operated by Akt . ^^^ This effect is dependent upon eNOS activation through an Akt dependent phosphorylation , as demonstrated by pharmacological modulation with l nitroarginine methyl ester and wortmannin and by Western blot studies . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In our present study , we investigated the effect of arsenite on Akt 1 and eNOS and its involvement in cytotoxicity of vascular endothelial cells . ^^^ Our study demonstrated that arsenite decreased the protein levels of both Akt 1 and eNOS accompanied with increased levels of ubiquitination of total cell lysates . ^^^ We found that inhibition of the ubiquitin proteasome pathway by MG 132 could partially protect Akt 1 and eNOS from degradation by arsenite together with a proportional protection from the arsenite induced cytoxicity . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
RNAi mediated down regulation of endogenous p66shc led to activation of the proto oncogene ras , and Akt kinase , with a corresponding increase in phosphorylation of eNOS at S 1177 ( S 1179 on bovine eNOS ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Despite the narrowing of arterial diameter and reduced expression of eNOS , expressions of phosphorylated protein kinase B ( Akt ) and phosphorylated eNOS were significantly increased in spastic arteries . ^^^ Transduction with AdEpo further increased the expression of phosphorylated Akt and eNOS and elevated basal levels of cGMP in the spastic arteries . ^^^ The vascular protective effect of Epo against cerebral vasospasm induced by SAH may be mediated in part by phosphorylation of Akt / eNOS . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
METHODS AND RESULTS : In human endothelial cells , we found that eNOS activation by TNF alpha is time dependent and requires activation of Akt , a known eNOS activator . eNOS activation was preceded by sequential activation of neutral sphingomyelinase 2 ( N SMase 2 ) and sphingosine kinase 1 ( SK 1 ) and generation of sphingosine 1 phosphate ( Sph1P ) . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Fluid shear stress elicited the phosphorylation and activation of Akt and eNOS as well as the tyrosine phosphorylation of Gab 1 and its association with the p 85 subunit of phosphatidylinositol 3 kinase and SHP 2 . ^^^ Overexpression of a Gab 1 mutant lacking the pleckstrin homology domain abrogated the shear stress induced phosphorylation of Akt but failed to affect the phosphorylation or activity of eNOS . ^^^ Mutation of Gab 1 Tyr627 to phenylalanine ( YF Gab 1 ) to prevent the binding of SHP 2 completely prevented the shear stress induced phosphorylation of eNOS , leaving the Akt response intact . ^^^ A dominant negative SHP 2 mutant prevented the activation of PKA and phosphorylation of eNOS without affecting that of Akt . ^^^ Thus , the shear stress induced activation of eNOS depends on Gab 1 and SHP 2 , which , in turn , regulate the phosphorylation and activity of eNOS by a PKA dependent but Akt independent mechanism . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This was accompanied by significant glomerulosclerosis , tubulointerstitial damage , renal immune cell infiltration , marked down regulations of renal tissue eNOS and nNOS , mild reduction of caveolin 1 , and unchanged calmodulin , phospho AKt , and sGC . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Human cytomegalovirus inhibits Akt mediated eNOS activation through upregulating PTEN ( phosphatase and tensin homolog deleted on chromosome 10 ) . ^^^ The signaling upstream of eNOS involving Akt and PDK 1 were also suppressed by the HCMV infection . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The transcription of eNOS induced by sesamol was confirmed through the activation of PI 3 kinase Akt ( protein kinase B ) signaling . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Contribution of the phosphatidylinositol 3 kinase / serine / threonine kinase ( PI3k / Akt ) pathway was assessed by measuring total and phosphorylated PI3K , Akt , BAD , and endothelial nitric oxide synthase ( eNOS ) proteins . ^^^ There was no significant induction of Akt activity or changes in phosphorylated BAD , Akt , or eNOS levels . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
PGE 2 increased phosphorylation of Akt and endothelial nitric oxide synthase ( eNOS ) , eNOS activity , and nitric oxide ( NO ) production by the activation of cAMP dependent protein kinase ( PKA ) and phosphatidylinositol 3 kinase ( PI3K ) . ^^^ These results clearly show that PGE 2 increased angiogenesis by activating the NO / cGMP signaling pathway through PKA / PI3K / Akt dependent increase in eNOS activity . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The FGF 2 mechanism responsible for reversing damages , involves the downstream enhancement of Akt , a pathway independent of eNOS activation . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
VEGF signaling leads to the phosphorylation of Akt and endothelial nitric oxide synthase ( p Akt and p eNOS ) . ^^^ The reduction in p Akt / Akt with the cholesterol diet at 2 weeks was not significantly different , but the correlation between the duration of cholesterol feeding and the reduction in p Akt / Akt was high ( r ( 2 ) = 0 . 858 ) . eNOS protein or mRNA did not change with cholesterol feeding , but p eNOS was significantly decreased at 4 . 5 weeks and all subsequent time points . nNOS mRNA and protein levels were decreased at 4 . 5 weeks and all subsequent time points , while iNOS was not different between groups . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
AMP signal transduction induces eNOS activation by promoting PKB phosphorylation . ^^^ Forskolin ( an activator of adenylyl cyclase that increases intracellular cAMP level ) and 8 bromo cAMP ( a membrane permeable cAMP analog ) were used to stimulate NO release and activation of PKB and endothelial NO synthase ( eNOS ) in these blood vessels . ^^^ N ( omega ) nitro l arginine methyl ester ( an NOS inhibitor ) blocked NO formation , Rp diastereomer of cAMP ( a PKA inhibitor ) , and LY 294002 [ a PI 3 kinase ( an activator of PKB ) inhibitor ] prevented the production of NO , phosphorylation of PKB , and eNOS induced by forskolin . ^^^ We are reporting for the first time that cAMP signal transduction stimulates eNOS activation through a PKB mediated mechanism . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
No change in ATP stimulated ERK 2 , Akt , or eNOS phosphorylation is observed with U 73122 ( 0 . 01 1 microM ) or 2 APB ( 1 50 microM ) . ^^^ Expression of constitutively active Akt ( ca Akt ) in UAEC resulted in slight elevation of basal eNOS activity , but relative ATP stimulated eNOS activation was not altered by ca Akt . ^^^ Wortmannin continued to inhibit eNOS activation by ATP in the presence of ca Akt ; LY 294002 still had no inhibitory effect . ^^^ We report that pregnancy adaptation of eNOS activation includes the reduced sensitivity to ERK mediated attenuation of eNOS activity and enhanced stimulation of eNOS activity through a wortmannin sensitive , LY 294002 insensitive , Akt independent mechanism . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Correspondingly , the activities of endothelial nitric oxide synthase ( eNOS ) and Akt / PKB ( measured by phosphorylation at serine 1 , 177 and serine 473 , respectively ) , showed two peaks over time . ^^^ Application of Gd ( 3+ ) , a potent SA channel blocker , and depletion of external Ca ( 2+ ) exclusively inhibited the first peaks of eNOS and Akt activity , but exerted little effect on the second peak . ^^^ These results suggest that up regulation of eNOS in response to cyclic stretch was mediated by two distinct pathways , [ Ca ( 2+ ) ] ( 1 ) increases via the SA channel in an early phase ( partially Akt / PKB ) , and PI3K Akt / PKB pathways in a late phase . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
To elucidate the beneficial cardioprotective mechanism of eplerenone , a novel selective aldosterone blocker , we hypothesized that eplerenone stimulates endothelial NO synthase ( eNOS ) through Akt and inhibits inducible NO synthase ( iNOS ) via nuclear factor kappaB ( NF kappaB ) after the development of oxidative stress and activation of the lectin like , oxidized , low density lipoprotein receptor 1 ( LOX 1 ) pathway in Dahl salt sensitive rats with heart failure . ^^^ Downregulated eNOS expression , eNOS and Akt phosphorylation , and NOS activity in failing rats were increased by eplerenone . ^^^ These findings suggest that eplerenone may have significant therapeutic potential for heart failure , and these cardioprotective mechanisms of eplerenone may be mediated in part by stimulating eNOS through Akt and inhibiting iNOS via NF kappaB after activation of the oxidative stress LOX 1 pathway and signal transduction pathway . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In endothelial cell culture , overexpression of PKCbeta 1 and beta 2 , but not PKCalpha , delta , or zeta , decreased insulin stimulated Akt phosphorylation and eNOS expression . ^^^ Thus , activation of PKCbeta in endothelial cells and vascular tissue inhibits Akt activation by insulin and VEGF , inhibits Akt dependent eNOS regulation by insulin , and causes endothelial dysfunction in obesity associated insulin resistance . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Statin therapy enhanced bone marrow VEGF protein levels , Akt phosphorylation , eNOS activity and normalized increased ROS levels . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
With specific inhibitors ( wortmannin and LY 294002 ) , we found that PI3K / Akt signaling participated in the eNOS phosphorylation caused by TNT , whereas the ERK pathway did not . ^^^ However , TNT mediated phosphorylation of either eNOS or Akt was drastically blocked by NAC and PEG CAT . ^^^ Interestingly , pretreatment with apocynin , a specific inhibitor for NADPH oxidase , diminished the phosphorylation of eNOS and Akt . ^^^ These results suggest that TNT affects NADPH oxidase , thereby generating hydrogen peroxide , which is capable of activating PI3K / Akt signaling associated with eNOS Ser 1179 phosphorylation . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Western blot analysis revealed that OxLDL dose dependently decreased Akt phosphorylation and eNOS protein expression and increased LOX 1 protein expression . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
This occurred via an Arg Gly Asp ( RGD ) peptide independent pathway through activation of G ( i / o ) proteins , phosphatidylinositol 3 kinase , Akt , and eNOS . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Phosphorylation of eNOS at Ser 617 ( bovine sequence ) is thought to occur in response to Akt stimulation and to be required for eNOS activity . ^^^ Together with basal eNOS activity , eNOS phosphorylation at Ser 617 and Akt Ser 473 phosphorylation were dose dependently and concomitantly suppressed by QRN within 30 min . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Vascular endothelial growth factor ( VEGF ) is implicated in angiogenesis , which activated angiogenic signaling cascade through Akt and endothelial nitric oxide synthase ( eNOS ) related pathway . ^^^ The phosphorylation of Akt protein and eNOS protein in the heart corresponded to the changes in the VEGF protein levels . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
We treated bovine pulmonary artery endothelial cells ( BPAECs ) with 20 HETE ( 1 microM ) or VEGF ( 8 . 3 nM ) and examined three molecular events known to activate eNOS : 1 ) phosphorylation at serine 1179 , 2 ) phosphorylation of protein kinase B ( Akt ) , which subsequently phosphorylates eNOS , and 3 ) association of eNOS with 90 kDa heat shock protein ( Hsp 90 ) . ^^^ Both 20 HETE and VEGF increase the phosphorylation of eNOS at serine 1179 and Akt at serine 473 . ^^^ In conclusion , VEGF and 20 HETE share eNOS activation pathways , including phosphorylation of serine 1179 and phosphorylation of Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In in vitro experiments , cell survival factors such as Akt and eNOS were significantly increased in BMSCs following anoxia . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
In particular , phosphorylation of serine 1179 ( bovine ) / 1177 ( human ) by Akt has been shown to be the central mechanism of eNOS regulation . ^^^ Proteasomal inhibition markedly enhanced PP2A association to eNOS , and this increase of PP2A dephosphorylated eNOS and its upstream kinase Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Analysis by RT PCR and Western blot showed significantly lower gene expression of vascular endothelial growth factor ( VEGF ) , angiopoietin 1 , and angiopoietin 2 and significantly lower protein expression of VEGF , angiopoietin 1 , angiopoietin 2 , the ratio of phospho Akt to Akt , and phospho endothelial nitric oxide synthase ( eNOS ) to eNOS in hypercholesterolemic rats than in controls . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Vascular endothelial growth factor induced eNOS Ser 1177 but not Akt Ser 473 phosphorylation levels were reduced significantly by pretreatment with Aldo . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Rapid phosphorylation of ERK1 / 2 , protein kinase B / Akt , and eNOS serine 1177 by equol was paralleled by association of eNOS with heat shock protein 90 ( Hsp 90 ) and NO synthesis in human umbilical vein endothelial cells , expressing estrogen receptors ( ER ) alpha and ERbeta . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Administration of recombinant AM was associated with augmented phosphorylation of Akt and eNOS in early reperfusion suggesting a role for AM in regulating this survival pathway . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
While linoleic acid inhibited Akt mediated eNOS phosphorylation , palmitic acid appeared to affect the upstream signaling . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Also , in 2 different models of active systemic anaphylaxis , inhibition of NOS , PI3K , or Akt or eNOS deficiency provided complete protection . ^^^ In contrast to the unsubstantiated paradigm that only excessive iNOS derived NO underlies cardiovascular collapse in shock , our data strongly support the unexpected concept that eNOS derived NO is the principal vasodilator in anaphylactic shock and define eNOS and / or PI3K or Akt as new potential targets for treating anaphylaxis . . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
MAIN OUTCOME MEASURE ( S ) : Measure of NO release , endothelial nitric oxide synthase ( eNOS ) activity and expression , and activation of ERK 1 / 2 and Akt . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Src activation induced the phosphorylation of caveolin 1 , Akt and eNOS , and promoted dissociation of eNOS from caveolin 1 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The receptor for PAF coupled to G ( 1 ) activates PI3Kgamma , which , in turn , is essential to promote Akt phosphorylation , NOSIII ( nitric oxide synthase isoform 3 ) activation and the production of nitric oxide , a well characterized cardiodepressing agent . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Since Ca ( 2+ ) independent NOS 3 activation may occur through its serine phosphorylation via protein kinase A ( PKA ) or Akt , we determined the PKA and Akt dependency of beta ( 2 ) adrenergic relaxation of rat aorta . ^^^ Western blotting of NOS 3 immunoprecipitates from rat aortas confirmed that albuterol increased serine phosphorylation of NOS 3 , and this increase was attenuated by H 89 or Akt inhibitor . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
Immunoblot analysis revealed that testosterone induced phosphorylation of Akt and NOS 3 . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
The expression of Nos 3 and Casp 3 was evaluated by real time PCR , and the activation state of PKB / Akt was assessed by western blot analysis and immunohistochemistry . ^^^ In addition , benfotiamine prevented the vascular accumulation of advanced glycation end products and the induction of pro apoptotic caspase 3 , while restoring proper expression of Nos 3 and Akt in ischaemic muscles . ^^^
Interacting proteins: P29474 and P31749 Pubmed SVM Score :0.0
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