C-reactive protein adversely alters the protein-protein interaction of the endothelial isoform of nitric oxide synthase

Simona Valleggi, Sridevi Devaraj, Mohan R. Dasu, Ishwarlal Jialal

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

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. METHODS: We incubated HAECs with CRP (0, 12.5, and 25 mg/L) for 1, 6, or 24 h and assessed the interaction of these proteins with eNOS by immunoprecipitation and western blotting. RESULTS: CRP treatment (12.5 and 25 mg/L) of HAECs for 24 h significantly increased eNOS binding to caveolin-1 (40% and 54% increase, respectively; P < 0.05) and decreased binding to Hsp90 (33% and 66% decrease, respectively; P < 0.05). CRP (25 mg/L) also significantly decreased the binding of porin to eNOS (11% decrease, P < 0.05). Similar results were seen when HAECs were treated with CRP for 6 h. CONCLUSIONS: These negative protein-protein interactions of eNOS were able to partly explain the CRP-induced decreases in the activity of this critical enzyme, which caused endothelial dysfunction.

Original languageEnglish (US)
Pages (from-to)1345-1348
Number of pages4
JournalClinical Chemistry
Volume56
Issue number8
DOIs
StatePublished - Aug 2010

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Nitric Oxide Synthase Type III
Nitric Oxide Synthase
C-Reactive Protein
Protein Isoforms
Endothelial cells
HSP90 Heat-Shock Proteins
Caveolin 1
Porins
Endothelial Cells
Proteins
Enzymes
Immunoprecipitation
Western Blotting

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

C-reactive protein adversely alters the protein-protein interaction of the endothelial isoform of nitric oxide synthase. / Valleggi, Simona; Devaraj, Sridevi; Dasu, Mohan R.; Jialal, Ishwarlal.

In: Clinical Chemistry, Vol. 56, No. 8, 08.2010, p. 1345-1348.

Research output: Contribution to journalArticle

Valleggi, Simona ; Devaraj, Sridevi ; Dasu, Mohan R. ; Jialal, Ishwarlal. / C-reactive protein adversely alters the protein-protein interaction of the endothelial isoform of nitric oxide synthase. In: Clinical Chemistry. 2010 ; Vol. 56, No. 8. pp. 1345-1348.
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abstract = "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. METHODS: We incubated HAECs with CRP (0, 12.5, and 25 mg/L) for 1, 6, or 24 h and assessed the interaction of these proteins with eNOS by immunoprecipitation and western blotting. RESULTS: CRP treatment (12.5 and 25 mg/L) of HAECs for 24 h significantly increased eNOS binding to caveolin-1 (40{\%} and 54{\%} increase, respectively; P < 0.05) and decreased binding to Hsp90 (33{\%} and 66{\%} decrease, respectively; P < 0.05). CRP (25 mg/L) also significantly decreased the binding of porin to eNOS (11{\%} decrease, P < 0.05). Similar results were seen when HAECs were treated with CRP for 6 h. CONCLUSIONS: These negative protein-protein interactions of eNOS were able to partly explain the CRP-induced decreases in the activity of this critical enzyme, which caused endothelial dysfunction.",
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