Direct interaction of endothelial nitric oxide synthase and caveolin-1 inhibits synthase activity

Richard C Venema, H. Ju, R. Zou, V. J. Venema

Research output: Contribution to journalArticle

Abstract

Endothelial nitric oxide synthase (eNOS) generates NO from L-arginine fol: lowing activation of the enzyme by Ca2+/calmodulin. NO synthesized by eNOS plays a key role in regulation of vascular tone and platelet aggregation. Association of eNOS with caveolin-1 in plasmalemmal caveolae has been shown previously in coimmunoprecipitation experiments. It is not known, however, whether eNOS and caveolin-1 interact directly or indirectly or whether interaction affects eNOS activity, eNOS contains an N-terminal oxygenase domain and a C-terminal reductase domain. Caveolin-1 Contains three domains consisting of N- and C-terminal cytosolic domains separated in the polypeptide sequence by a membrane spanning domain. To determine whether eNOS and caveolin-1 interact directly and to map the interacting domains in the two proteins, we have investigated the eNOS-caveolin-1 interaction using an in vitro binding assay with glutathione S-transferase fusion proteins. We have also mapped the domains involved in the interaction using a yeast two-hybrid system. Results obtained using both approaches show that both N- and Cterminal cytosolic domains of caveolin-1 interact directly with the eNOS oxygenase domain. Furthermore, interaction of eNOS with caveolin-1 significantly inhibits eNOS catalytic activity. Negative allosteric regulation of eNOS by caveolin-1 may thus function in opposition to the well-known positive allosteric regulation of eNOS by Ca2+/calmodulin.

Original languageEnglish (US)
JournalFASEB Journal
Volume11
Issue number9
StatePublished - Dec 1 1997

Fingerprint

Caveolin 1
Nitric Oxide Synthase Type III
Allosteric Regulation
Oxygenases
calmodulin
Calmodulin
oxygenases
endothelial nitric oxide synthase
calcium
Caveolae
two hybrid system techniques
enzyme activation
Two-Hybrid System Techniques
Enzyme Activation
platelet aggregation
Platelets
Glutathione Transferase
Hybrid systems
Platelet Aggregation
catalytic activity

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Direct interaction of endothelial nitric oxide synthase and caveolin-1 inhibits synthase activity. / Venema, Richard C; Ju, H.; Zou, R.; Venema, V. J.

In: FASEB Journal, Vol. 11, No. 9, 01.12.1997.

Research output: Contribution to journalArticle

Venema, Richard C ; Ju, H. ; Zou, R. ; Venema, V. J. / Direct interaction of endothelial nitric oxide synthase and caveolin-1 inhibits synthase activity. In: FASEB Journal. 1997 ; Vol. 11, No. 9.
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