Dehydroepiandrosterone activates endothelial cell nitric-oxide synthase by a specific plasma membrane receptor coupled to Galpha(i2,3)

Dongmin Liu; Joseph S Dillon

doi:10.1074/jbc.M200491200

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Dehydroepiandrosterone activates endothelial cell nitric-oxide synthase by a specific plasma membrane receptor coupled to Galpha(i2,3)

Journal article

Open access

Peer reviewed

Dehydroepiandrosterone activates endothelial cell nitric-oxide synthase by a specific plasma membrane receptor coupled to Galpha(i2,3)

Dongmin Liu and Joseph S Dillon

The Journal of biological chemistry, Vol.277(24), pp.21379-21388

06/14/2002

DOI: 10.1074/jbc.M200491200

PMID: 11934890

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Abstract

The adrenal steroid dehydroepiandrosterone (DHEA) has no known cellular receptor or unifying mechanism of action, despite evidence suggesting beneficial vascular effects in humans. Based on previous data from our laboratory, we hypothesized that DHEA binds to specific cell-surface receptors to activate intracellular G-proteins and endothelial nitric-oxide synthase (eNOS). We now pharmacologically characterize a putative plasma membrane DHEA receptor and define its associated G-proteins. The [3H]DHEA binding to isolated plasma membranes from bovine aortic endothelial cells was of high affinity (K(d) = 48.7 pm) and saturable (B(max) = 500 fmol/mg protein). Structurally related steroids failed to compete with DHEA for binding. The putative DHEA receptor was functionally coupled to G-proteins, because guanosine 5'-O-(3-thio)triphosphate (GTPgammaS) inhibited [3H]DHEA binding to plasma membranes by 69%, and DHEA increased [35S]GTPgammaS binding by 157%. DHEA stimulated [35S]GTPgammaS binding to Galpha(i2) and Galpha(i3), but not to Galpha(i1) or Galpha(o). Pretreatment of plasma membranes with antibody to Galpha(i2) or Galpha(i3), but not to Galpha(i1), inhibited the DHEA activation of eNOS. Thus, DHEA receptors are expressed on endothelial cell plasma membranes and are coupled to eNOS activity through Galpha(i2) and Galpha(i3). These novel findings should allow us to isolate the putative receptor and reevaluate the physiological role of DHEA activity.

Adjuvants, Immunologic - pharmacology

GTP-Binding Proteins - chemistry

Dehydroepiandrosterone - pharmacology

Receptors, Estrogen - antagonists & inhibitors

Virulence Factors, Bordetella - pharmacology

Endothelium, Vascular - enzymology

Blotting, Western

Precipitin Tests

Tissue Distribution

Guanosine 5'-O-(3-Thiotriphosphate) - metabolism

Animals

Time Factors

Cattle

Protein Binding

Steroids - pharmacology

Cell Membrane - metabolism

Nitric Oxide Synthase - metabolism

Enzyme Activation

Kinetics

Aorta - enzymology

GTP-Binding Proteins - metabolism

Details

Title: Subtitle: Dehydroepiandrosterone activates endothelial cell nitric-oxide synthase by a specific plasma membrane receptor coupled to Galpha(i2,3)
Creators: Dongmin Liu - Division of Endocrinology, Department of Internal Medicine, University of Iowa College of Medicine and Veterans Administration Medical Center, Iowa City, Iowa 52246, USA
Joseph S Dillon
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.277(24), pp.21379-21388
DOI: 10.1074/jbc.M200491200
PMID: 11934890
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Grant note: AG55741 / NIA NIH HHS R01 AG018928 / NIA NIH HHS DK25295 / NIDDK NIH HHS
Language: English
Date published: 06/14/2002
Academic Unit: Fraternal Order of Eagles Diabetes Research Center; Endocrinology and Metabolism; Internal Medicine
Record Identifier: 9984094740902771

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