Functional interaction of endothelial nitric oxide synthase with a voltage-dependent anion channel

Sun, Jianxin; Liao, James K.

doi:10.1073/pnas.202260999

Cited by 63 publications

(56 citation statements)

References 40 publications

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“…34,35 Immunoblotting was performed using monoclonal antibodies to eNOS (1:1000 dilution; Transduction Laboratories, Lexington, Ky), to p85 antiserum (1:500 dilution; Upstate Biotech), to phospho-eNOS (Ser1177), phospho-Akt (Ser473), and Akt (1:1000 dilution; Cell Signaling Tech), to hsp90 (1:200 dilution; Santa Cruz Biotech), and to ␣ -tubulin (DM1A) (1:5000 dilution; Sigma). Immunodetection was accomplished using a sheep anti-mouse secondary antibody (1:2000 dilution) or donkey anti-rabbit secondary antibody (1:2000 dilution) and the enhanced chemiluminescence (ECL) kit (Amersham Corp).…”

Section: Western Blotting and Immunoprecipitationmentioning

confidence: 99%

Induction of Angiogenesis by Heat Shock Protein 90 Mediated by Protein Kinase Akt and Endothelial Nitric Oxide Synthase

Sun

Liao

2004

ATVB

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Objective-A specific inhibitor of heat shock protein 90 (Hsp90), 17-AAG, has been shown to inhibit tumor growth through cell cycle arrest, differentiation, or apoptosis. Because angiogenesis is important for tumor growth, we hypothesize that inhibition of angiogenesis by 17-AAG may mediate some of its antitumor effects. 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 Hsp90 by 17-AAG decreased Akt and eNOS expression by 74% and 81%, respectively. Inhibition of eNOS expression by 17-AAG occurred at the transcriptional level as determined by eNOS promoter activity and nuclear run-on assay. Furthermore, treatment with 17-AAG decreased basal and vascular endothelial growth factor-stimulated Akt and eNOS phosphorylation. This corresponded with decreased NO production and inhibition of endothelial cell migration and angiogenesis. The anti-angiogenic effect of 17-AAG was partially reversed by the NO donor, SNAP. Conclusions-These

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Section: Western Blotting and Immunoprecipitationmentioning

confidence: 99%

Induction of Angiogenesis by Heat Shock Protein 90 Mediated by Protein Kinase Akt and Endothelial Nitric Oxide Synthase

Sun

Liao

2004

ATVB

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“…Furthermore, numerous groups have shown that CRP impairs endothelial vasoreactivity in vivo (11 ). To date, eNOS has been shown to interact directly with several proteins, including heat shock protein 90 (Hsp90), calmodulin, dynamin-2, caveolin-1, the intracellular domains of certain G protein-coupled receptors, and porin, a voltage-dependent anion/cation channel (5)(6)(7)(12)(13)(14)(15). The most important interactions for activity appear to relate to its association with caveolin-1 (decreased activity) and Hsp90 (increased activity) (5)(6)(7).…”

Section: Discussionmentioning

confidence: 99%

C-Reactive Protein Adversely Alters the Protein–Protein Interaction of the Endothelial Isoform of Nitric Oxide Synthase

et al. 2010

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BACKGROUND: C-reactive protein (CRP) inhibits the activity of the endothelial isoform of nitric oxide synthase (eNOS) via uncoupling of the enzyme both in vitro and in vivo. eNOS activity appears to be related in part to its interaction with other cellular proteins, including heat shock protein 90 (Hsp90), caveolin-1, and porin. In this study, we examined the effect of CRP treatment of human aortic endothelial cells (HAECs) on eNOS interaction with caveolin-1, Hsp90, and porin.

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“…However, caveolin, which competes with and displaces calmodulin from eNOS, inhibits eNOS activity in the caveolae of endothelial cells and cardiomyocytes (10,11). Other eNOS interacting proteins such as heat shock protein 90 and a voltage-dependent anion/cation channel, porin, also regulate eNOS activation through protein stabilization, sub-cellular localization, and cofactor availability (12,13). Thus, proteins that interact with eNOS are important physiological regulators of eNOS activity.…”

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confidence: 99%

Dynamic regulation of endothelial NOS mediated by competitive interaction with α‐actinin‐4 and calmodulin

Hiroi

Guo

et al. 2008

FASEB j.

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Alpha-actinins are critical components of the actin cytoskeleton. Here we show that α-actinins serve another important biological function by binding to and competitively inhibiting calcium-dependent activation of endothelial NOS (eNOS). α-actinin-2 was found to associate with eNOS in a yeast twohybrid screen. In vascular endothelial cells, which only express α-actinin-1 and -4, α-actinin-4 interacted and colocalized with eNOS. Addition of α-actinin-4 directly inhibited eNOS recombinant protein, and overexpression of α-actinin-4 inhibited eNOS activity in eNOS-transfected COS-7 cells and bovine aortic endothelial cells (BAECs). In contrast, knockdown of α-actinin-4 by siRNA increased eNOS activity in BAECs. The α-actinin-4-binding site on eNOS was mapped to a central region comprising the calmodulin-binding domain, and the eNOS-binding site on α-actinin-4 was mapped to the fourth spectrin-like rod domain, R4. Treatment of endothelial cells with a calcium ionophore, A23187, decreased α-actinin-4-eNOS interaction, leading to translocation of α-actinin-4 from plasma membrane to cytoplasm. Indeed, addition of calmodulin displaced α-actinin-4 binding to eNOS and increased eNOS activity. These findings indicate that eNOS activity in vascular endothelial cells is tonically and dynamically regulated by competitive interaction with α-actinin-4 and calmodulin. Keywords endothelium; nitric oxide; ionophore Endothelium-derived NO is an important mediator of vascular function (1). NO is generated from the conversion of L-arginine to L-citrulline by endothelial NOS (eNOS or NOS3). Because of its critical role in vascular homeostasis, eNOS is tightly regulated posttranslationally by several factors. Lipid modifications such as myristoylation or palmitoylation are important to anchor eNOS at the cell membrane, in close proximity to its cofactors and modulators (2). Upstream modulators include protein kinases, which activate eNOS through phosphorylation at Ser-1177 and Ser-635 or inhibit eNOS through phosphorylation at Thr-495 and Ser-116 3 . In particular, Ser-1177 phosphorylation of eNOS by protein kinase B/Akt enhances both basal and agonist-mediated eNOS activity (4,5). In contrast, S-nitrosylation of eNOS renders eNOS inactive under basal conditions (6,7).

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Functional interaction of endothelial nitric oxide synthase with a voltage-dependent anion channel

Cited by 63 publications

References 40 publications

Induction of Angiogenesis by Heat Shock Protein 90 Mediated by Protein Kinase Akt and Endothelial Nitric Oxide Synthase

Induction of Angiogenesis by Heat Shock Protein 90 Mediated by Protein Kinase Akt and Endothelial Nitric Oxide Synthase

C-Reactive Protein Adversely Alters the Protein–Protein Interaction of the Endothelial Isoform of Nitric Oxide Synthase

Dynamic regulation of endothelial NOS mediated by competitive interaction with α‐actinin‐4 and calmodulin

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