Endothelin receptor is coupled to phospholipase C via a pertussis toxin-insensitive guanine nucleotide-binding regulatory protein in vascular smooth muscle cells

Yoh Takuwa, Yoshitoshi Kasuya, Noriko Takuwa, Michiyo Kudo, Masashi Yanagisawa, Katsutoshi Goto, Tomoh Masaki, Kamejiro Yamashita

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

The mechanisms of endothelin-1 (ET) actions were investigated in cultured rat aortic vascular smooth muscle A-10 cells. The A-10 cells have a single class of high affinity binding sites for ET with an apparent Mr of 65,000-75,000 on SDS-PAGE. Stimulation of cells with ET induces mobilization of Ca2+ from both intra- and extracellular pools to produce a bip basic increase in cytoplasmic free Ca2+ concentration. ET increases cellular levels of inositol trisphosphate and 1,2-diacylglycerol, indicating activation of phospholipase C by ET. ET stimulates production of inositol phosphates in membranes prepared from A-10 cells in the presence of guanosine 5′-O-(thiotriphosphate) (GTPγS), but not in its absence. Further, specific binding of 125I-labeled ET to A-10 cell membranes is shown to be inhibited by GTPγS in a dose-dependent manner. Treatment of A-10 cells with pertussis toxin induces ADP-ribosylation of a 41,000-D membrane protein but fails to block the ET-induced increases in inositol phosphate production and Ca2+ mobilization. These results indicate that the receptor for ET is coupled to phospholipase C via a guanine nucleotide-binding regulatory protein which is distinct from the pertussis toxin substrate in A-10 cells.

Original languageEnglish (US)
Pages (from-to)653-658
Number of pages6
JournalJournal of Clinical Investigation
Volume85
Issue number3
StatePublished - Mar 1990

Fingerprint

Endothelin Receptors
Pertussis Toxin
Type C Phospholipases
Endothelin-1
Vascular Smooth Muscle
GTP-Binding Proteins
Smooth Muscle Myocytes
antineoplaston A10
Carrier Proteins
Inositol Phosphates
Endothelin A Receptors
Guanosine
Inositol
Adenosine Diphosphate
Polyacrylamide Gel Electrophoresis
Membrane Proteins
Binding Sites
Cell Membrane
Membranes

Keywords

  • 1,2-diacylglycerol
  • Ca channel
  • Inositol phosphate
  • Phosphoinositide hydrolysis
  • Vasoconstriction

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Endothelin receptor is coupled to phospholipase C via a pertussis toxin-insensitive guanine nucleotide-binding regulatory protein in vascular smooth muscle cells. / Takuwa, Yoh; Kasuya, Yoshitoshi; Takuwa, Noriko; Kudo, Michiyo; Yanagisawa, Masashi; Goto, Katsutoshi; Masaki, Tomoh; Yamashita, Kamejiro.

In: Journal of Clinical Investigation, Vol. 85, No. 3, 03.1990, p. 653-658.

Research output: Contribution to journalArticle

Takuwa, Yoh ; Kasuya, Yoshitoshi ; Takuwa, Noriko ; Kudo, Michiyo ; Yanagisawa, Masashi ; Goto, Katsutoshi ; Masaki, Tomoh ; Yamashita, Kamejiro. / Endothelin receptor is coupled to phospholipase C via a pertussis toxin-insensitive guanine nucleotide-binding regulatory protein in vascular smooth muscle cells. In: Journal of Clinical Investigation. 1990 ; Vol. 85, No. 3. pp. 653-658.
@article{77a44bdbc9d9477eaee3fc783e15fd65,
title = "Endothelin receptor is coupled to phospholipase C via a pertussis toxin-insensitive guanine nucleotide-binding regulatory protein in vascular smooth muscle cells",
abstract = "The mechanisms of endothelin-1 (ET) actions were investigated in cultured rat aortic vascular smooth muscle A-10 cells. The A-10 cells have a single class of high affinity binding sites for ET with an apparent Mr of 65,000-75,000 on SDS-PAGE. Stimulation of cells with ET induces mobilization of Ca2+ from both intra- and extracellular pools to produce a bip basic increase in cytoplasmic free Ca2+ concentration. ET increases cellular levels of inositol trisphosphate and 1,2-diacylglycerol, indicating activation of phospholipase C by ET. ET stimulates production of inositol phosphates in membranes prepared from A-10 cells in the presence of guanosine 5′-O-(thiotriphosphate) (GTPγS), but not in its absence. Further, specific binding of 125I-labeled ET to A-10 cell membranes is shown to be inhibited by GTPγS in a dose-dependent manner. Treatment of A-10 cells with pertussis toxin induces ADP-ribosylation of a 41,000-D membrane protein but fails to block the ET-induced increases in inositol phosphate production and Ca2+ mobilization. These results indicate that the receptor for ET is coupled to phospholipase C via a guanine nucleotide-binding regulatory protein which is distinct from the pertussis toxin substrate in A-10 cells.",
keywords = "1,2-diacylglycerol, Ca channel, Inositol phosphate, Phosphoinositide hydrolysis, Vasoconstriction",
author = "Yoh Takuwa and Yoshitoshi Kasuya and Noriko Takuwa and Michiyo Kudo and Masashi Yanagisawa and Katsutoshi Goto and Tomoh Masaki and Kamejiro Yamashita",
year = "1990",
month = "3",
language = "English (US)",
volume = "85",
pages = "653--658",
journal = "Journal of Clinical Investigation",
issn = "0021-9738",
publisher = "The American Society for Clinical Investigation",
number = "3",

}

TY - JOUR

T1 - Endothelin receptor is coupled to phospholipase C via a pertussis toxin-insensitive guanine nucleotide-binding regulatory protein in vascular smooth muscle cells

AU - Takuwa, Yoh

AU - Kasuya, Yoshitoshi

AU - Takuwa, Noriko

AU - Kudo, Michiyo

AU - Yanagisawa, Masashi

AU - Goto, Katsutoshi

AU - Masaki, Tomoh

AU - Yamashita, Kamejiro

PY - 1990/3

Y1 - 1990/3

N2 - The mechanisms of endothelin-1 (ET) actions were investigated in cultured rat aortic vascular smooth muscle A-10 cells. The A-10 cells have a single class of high affinity binding sites for ET with an apparent Mr of 65,000-75,000 on SDS-PAGE. Stimulation of cells with ET induces mobilization of Ca2+ from both intra- and extracellular pools to produce a bip basic increase in cytoplasmic free Ca2+ concentration. ET increases cellular levels of inositol trisphosphate and 1,2-diacylglycerol, indicating activation of phospholipase C by ET. ET stimulates production of inositol phosphates in membranes prepared from A-10 cells in the presence of guanosine 5′-O-(thiotriphosphate) (GTPγS), but not in its absence. Further, specific binding of 125I-labeled ET to A-10 cell membranes is shown to be inhibited by GTPγS in a dose-dependent manner. Treatment of A-10 cells with pertussis toxin induces ADP-ribosylation of a 41,000-D membrane protein but fails to block the ET-induced increases in inositol phosphate production and Ca2+ mobilization. These results indicate that the receptor for ET is coupled to phospholipase C via a guanine nucleotide-binding regulatory protein which is distinct from the pertussis toxin substrate in A-10 cells.

AB - The mechanisms of endothelin-1 (ET) actions were investigated in cultured rat aortic vascular smooth muscle A-10 cells. The A-10 cells have a single class of high affinity binding sites for ET with an apparent Mr of 65,000-75,000 on SDS-PAGE. Stimulation of cells with ET induces mobilization of Ca2+ from both intra- and extracellular pools to produce a bip basic increase in cytoplasmic free Ca2+ concentration. ET increases cellular levels of inositol trisphosphate and 1,2-diacylglycerol, indicating activation of phospholipase C by ET. ET stimulates production of inositol phosphates in membranes prepared from A-10 cells in the presence of guanosine 5′-O-(thiotriphosphate) (GTPγS), but not in its absence. Further, specific binding of 125I-labeled ET to A-10 cell membranes is shown to be inhibited by GTPγS in a dose-dependent manner. Treatment of A-10 cells with pertussis toxin induces ADP-ribosylation of a 41,000-D membrane protein but fails to block the ET-induced increases in inositol phosphate production and Ca2+ mobilization. These results indicate that the receptor for ET is coupled to phospholipase C via a guanine nucleotide-binding regulatory protein which is distinct from the pertussis toxin substrate in A-10 cells.

KW - 1,2-diacylglycerol

KW - Ca channel

KW - Inositol phosphate

KW - Phosphoinositide hydrolysis

KW - Vasoconstriction

UR - http://www.scopus.com/inward/record.url?scp=0025265191&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025265191&partnerID=8YFLogxK

M3 - Article

C2 - 2155922

AN - SCOPUS:0025265191

VL - 85

SP - 653

EP - 658

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

SN - 0021-9738

IS - 3

ER -