Endothelial nitric oxide synthase interactions with G-protein-coupled receptors

Mario B Marrero, Virginia J. Venema, Hong Ju, Han He, Haiying Liang, Ruth B Caldwell, Richard C Venema

Research output: Contribution to journalArticle

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Abstract

The endothelial nitric oxide synthase (eNOS) is activated in response to stimulation of endothelial cells by a number of vasoactive substances including, bradykinin (BK), angiotensin II (Ang II), endothelin-1 (ET-1) and ATP. In the present study we have used in vitro activity assays of purified eNOS and in vitro binding assays with glutathione S-transferase fusion proteins to show that the capacity to bind and inhibit eNOS is a common feature of membrane-proximal regions of intracellular domain 4 of the BK B2, the Ang II AT1 and the ET-1 ETB receptors, but not of the ATP P2Y2 receptor. Phosphorylation of serine or tyrosine residues in the eNOS-interacting region of the B2 receptor results in a loss of eNOS inhibition due to a decrease in the binding affinity of the receptor domain for the eNOS enzyme. Furthermore, the B2 receptor is transiently phosphorylated on tyrosine residues in cultured endothelial cells in response to BK stimulation. Phosphorylation occurs during the time in which eNOS transiently dissociates from the receptor accompanied by a transient increase in nitric oxide production. Taken together, these data support the hypotheses that eNOS is regulated in endothelial cells by reversible and inhibitory interactions with G-protein-coupled receptors and that these interactions can be modulated by receptor phosphorylation.

Original languageEnglish (US)
Pages (from-to)335-340
Number of pages6
JournalBiochemical Journal
Volume343
Issue number2
DOIs
StatePublished - Oct 15 1999

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Nitric Oxide Synthase Type III
G-Protein-Coupled Receptors
Phosphorylation
Endothelial cells
Bradykinin
Endothelial Cells
Endothelin-1
Angiotensin II
Tyrosine
Assays
Adenosine Triphosphate
Purinergic P2Y2 Receptors
Purinergic P2 Receptors
Endothelin A Receptors
Glutathione Transferase
Serine
Cultured Cells
Nitric Oxide
Fusion reactions
Cell Count

Keywords

  • ATP
  • Angiotensin II
  • Bradykinin
  • Endothelin-1
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Endothelial nitric oxide synthase interactions with G-protein-coupled receptors. / Marrero, Mario B; Venema, Virginia J.; Ju, Hong; He, Han; Liang, Haiying; Caldwell, Ruth B; Venema, Richard C.

In: Biochemical Journal, Vol. 343, No. 2, 15.10.1999, p. 335-340.

Research output: Contribution to journalArticle

Marrero, MB, Venema, VJ, Ju, H, He, H, Liang, H, Caldwell, RB & Venema, RC 1999, 'Endothelial nitric oxide synthase interactions with G-protein-coupled receptors', Biochemical Journal, vol. 343, no. 2, pp. 335-340. https://doi.org/10.1042/0264-6021:3430335
Marrero, Mario B ; Venema, Virginia J. ; Ju, Hong ; He, Han ; Liang, Haiying ; Caldwell, Ruth B ; Venema, Richard C. / Endothelial nitric oxide synthase interactions with G-protein-coupled receptors. In: Biochemical Journal. 1999 ; Vol. 343, No. 2. pp. 335-340.
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