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Role of Caveolin‐1 S‐Nitrosylation, Ubiquitination, and Degradation in Idiopathic Pulmonary Arterial Hypertension
Author(s) -
Bakhshi Farnaz R,
Bonini M,
Chen Z,
NieuwAmerongen GN,
Comhair S,
Erzurum S,
Minshall RD
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.878.7
Subject(s) - enos , ubiquitin , chemistry , s nitrosylation , caveolin 1 , phosphorylation , hyperactivation , oxidative stress , cav1.2 , nitric oxide , microbiology and biotechnology , cancer research , biochemistry , calcium , medicine , biology , voltage dependent calcium channel , nitric oxide synthase , cysteine , organic chemistry , gene , enzyme
Cav1 −/− mice develop pulmonary arterial hypertension (PAH) due to eNOS hyperactivation since inhibition or deletion of eNOS abrogates the PAH phenotype. Here, we tested the hypothesis that oxidative stress upon inflammatory stimuli can induce Cav1 degradation & that this is associated with PAH. We observed TNFα‐induced NO production and S‐nitrosylation (SNO) at Cav1 C156 increased Src activity and destabilized Cav1 oligomeric chains in human pulmonary endothelial cells (PECs), and that this could be blocked by eNOS & Src inhibitors. Cav1 SNO at C156 was associated with reduced Cav1 expression following prolonged (72 hr) stimulation with TNFα or the NO donor, DetaNONOate. Mass spectrometry revealed Cav‐1 ubiquitination (Ub) on K86, supporting the hypothesis that Cav1 SNO at C156 induces Src‐dependent Cav1 Y14 phosphorylation, destabilization of Cav1 oligomers, & Ub of K86 leading to Cav1 degradation. In transduced HEK cells, we observed decreased Cav1 oligomerization with the Cav1 C156S‐mutant, preservation of oligomers with Y14F, and abrogation of Ub & degradation with a K86R mutant compared to WT‐Cav1. We also observed decreased oligomerized & total Cav1 expression as well as eNOS hyperactivation in PECs from 2 patients with IPAH compared to control PECs. Thus, we show oxidative stress‐induced Cav1 SNO, phosphorylation, ubiquitination, & degradation is associated with idiopathic lung vascular disease.