Inhibitory interactions of the bradykinin B2 receptor with endothelial nitric-oxide synthase

Hong Ju, Virginia J. Venema, Mario B Marrero, Richard C Venema

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

It has been shown previously that the endothelial nitric-oxide synthase (eNOS) interacts reversibly with the plasmalemmal caveolae structural protein, caveolin-1. The eNOS-caveolin-1 interaction inhibits eNOS catalytic activity. In the present study, we show that eNOS also participates in reversible inhibitory interactions with the G protein-coupled bradykinin B2 receptor. eNOS and the B2 receptor are coimmunoprecipitated from endothelial cell lysates by antibodies directed against either of the two proteins. A glutathione S-transferase fusion protein containing intracellular domain 4 of the receptor is bound by purified recombinant eNOS in in vitro binding assays. The fusion protein selectively inhibits the activity of purified eNOS. A synthetic peptide corresponding to membrane-proximal residues 310- 334 in intracellular domain 4 also potently inhibits eNOS activity (IC50 < 1 μM). Treatment of cultured endothelial cells with bradykinin or Ca2+ ionophore promotes a rapid dissociation of the eNOS·B2 receptor complex. These data demonstrate that the bradykinin B2 receptor physically associates with eNOS in a ligand- and Ca2+dependent manner. Reversible and inhibitory membrane-docking interactions of eNOS, therefore, are not restricted to those with caveolin-1 but also occur with the bradykinin B2 receptor.

Original languageEnglish (US)
Pages (from-to)24025-24029
Number of pages5
JournalJournal of Biological Chemistry
Volume273
Issue number37
DOIs
StatePublished - Sep 11 1998

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Bradykinin B2 Receptors
Nitric Oxide Synthase Type III
Caveolin 1
Endothelial cells
Proteins
Fusion reactions
Endothelial Cells
Membranes
Caveolae
Ionophores
Bradykinin
Glutathione Transferase
GTP-Binding Proteins
Inhibitory Concentration 50
Cultured Cells
Assays
Catalyst activity

ASJC Scopus subject areas

  • Biochemistry

Cite this

Inhibitory interactions of the bradykinin B2 receptor with endothelial nitric-oxide synthase. / Ju, Hong; Venema, Virginia J.; Marrero, Mario B; Venema, Richard C.

In: Journal of Biological Chemistry, Vol. 273, No. 37, 11.09.1998, p. 24025-24029.

Research output: Contribution to journalArticle

Ju, Hong ; Venema, Virginia J. ; Marrero, Mario B ; Venema, Richard C. / Inhibitory interactions of the bradykinin B2 receptor with endothelial nitric-oxide synthase. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 37. pp. 24025-24029.
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