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C‐Reactive Protein Impairs eNOS protein‐protein interactions
Author(s) -
Valleggi Simona,
Devaraj Sridevi,
Jialal Ishwarlal
Publication year - 2010
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.24.1_supplement.750.7
Subject(s) - enos , hsp90 , porin , medicine , endocrinology , heat shock protein , chemistry , biology , nitric oxide , biochemistry , nitric oxide synthase , escherichia coli , bacterial outer membrane , gene
C Reactive Protein (CRP) is an acute‐phase protein and a cardiovascular risk marker. CRP levels are inversely correlated with endothelial vasoreactivity. CRP decreases eNOS expression and activity in vitro and in vivo . We have shown that CRP mediates eNOS uncoupling. In this study, we tested the effect of CRP on eNOS interaction and association with docking proteins, a fundamental step in regulating eNOS activity. Human Aortic Endothelial cells were incubated in absence and presence of CRP (25ug/ml for 18hrs) followed by stimulation with VEGF (100ng/mL for 0–20 minutes). Association between eNOS and Caveolin‐1 (Cav1), Dynamin II (DynII), Porin and Heat Shock Protein 90 (Hsp90) was assessed by immunoprecipitating cell lysates with eNOS followed by blotting for Hsp90, DynII, Porin and Cav1, using eNOS as loading control (n=3–5 separate experiments). CRP treatment results in a 87% decrease in eNOS‐Hsp90 association, 26% decrease in eNOS‐Porin association and 33% increase in eNOS‐Cav1 association. No significant changes have been observed in eNOS‐DynII association. Thus, we provide further evidence for CRP inducing endothelial dysfunction in HAECs via changes in eNOS association with docking proteins such as Hsp90, Cav1 and Porin.