Differences in the regulation of endothelin-1- and lysophosphatidic-acid-stimulated Ins(1,4,5)P3 formation in rat-1 fibroblasts

R Plevin, E E MacNulty, S Palmer, M J Wakelam

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Endothelin-1 (ET-1)- and lysophosphatidic acid (LPA)-stimulated PtdIns(4,5)P2 hydrolysis has been studied in Rat-1 fibroblasts. Although both agonists caused the dose-dependent accumulation of inositol phosphates, a number of differences were observed. LPA induced a transient increase in Ins(1,4,5)P3 mass which returned to basal levels within 90 s, whereas the response to ET-1 did not desensitize, with levels remaining at 3-4 times basal values for up to 15 min. Stimulated decreases in mass levels of PtdIns(4,5)P2 mirrored Ins(1,4,5)P3 formation for both agonists. Experiments with electropermeabilized cells demonstrated that the effects of both agonists are stimulated by a phospholipase C controlled by a guanine-nucleotide-binding regulatory protein; however, there are differences in the nature of these interactions. The inositol phosphate response to ET-1 is poorly potentiated by guanosine 5'-[gamma-thio]triphosphate (GTP[S]) and markedly inhibited by guanosine 5'-[beta-thio]diphosphate (GDP[S]), whereas that to LPA is potentiated by GTP[S] but is relatively insensitive to GDP[S]. In addition, LPA decreased the lag time for the onset of GTP[S]-stimulated [3H]InsP3 accumulation, whereas ET-1 was without effect. Phorbol 12-myristate 13-acetate treatment of the cells inhibited LPA-stimulated, but not ET-1-stimulated, inositol phosphate formation in both intact and permeabilized cells, suggesting that the site of protein kinase C-mediated phosphorylation may be blocked in ET-1-stimulated Rat-1 cells. The results indicate that the receptor-G-protein-phospholipase C interaction for the two agonists may not conform to the same model.

LanguageEnglish
Pages609-615
Number of pages7
JournalBiochemical journal
Volume280
Issue number3
Publication statusPublished - 15 Dec 1991

Fingerprint

Endothelin-1
Fibroblasts
Rats
Inositol Phosphates
Guanosine Triphosphate
Phosphatidylinositol 4,5-Diphosphate
Type C Phospholipases
GTP-Binding Proteins
Guanosine 5'-O-(3-Thiotriphosphate)
Phosphorylation
Guanine Nucleotides
Guanosine
Protein C
Protein Kinase C
lysophosphatidic acid
Hydrolysis
Carrier Proteins
Acetates
Cells
Proteins

Keywords

  • animals
  • cell membrane permeability
  • cells, cultured
  • endothelins
  • fibroblasts
  • GTP-binding proteins
  • guanosine 5'-O-(3-Thiotriphosphate)
  • inositol phosphates
  • lysophospholipids
  • phosphatidylinositols
  • phosphorylation
  • rats
  • tetradecanoylphorbol acetate
  • type C phospholipases

Cite this

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title = "Differences in the regulation of endothelin-1- and lysophosphatidic-acid-stimulated Ins(1,4,5)P3 formation in rat-1 fibroblasts",
abstract = "Endothelin-1 (ET-1)- and lysophosphatidic acid (LPA)-stimulated PtdIns(4,5)P2 hydrolysis has been studied in Rat-1 fibroblasts. Although both agonists caused the dose-dependent accumulation of inositol phosphates, a number of differences were observed. LPA induced a transient increase in Ins(1,4,5)P3 mass which returned to basal levels within 90 s, whereas the response to ET-1 did not desensitize, with levels remaining at 3-4 times basal values for up to 15 min. Stimulated decreases in mass levels of PtdIns(4,5)P2 mirrored Ins(1,4,5)P3 formation for both agonists. Experiments with electropermeabilized cells demonstrated that the effects of both agonists are stimulated by a phospholipase C controlled by a guanine-nucleotide-binding regulatory protein; however, there are differences in the nature of these interactions. The inositol phosphate response to ET-1 is poorly potentiated by guanosine 5'-[gamma-thio]triphosphate (GTP[S]) and markedly inhibited by guanosine 5'-[beta-thio]diphosphate (GDP[S]), whereas that to LPA is potentiated by GTP[S] but is relatively insensitive to GDP[S]. In addition, LPA decreased the lag time for the onset of GTP[S]-stimulated [3H]InsP3 accumulation, whereas ET-1 was without effect. Phorbol 12-myristate 13-acetate treatment of the cells inhibited LPA-stimulated, but not ET-1-stimulated, inositol phosphate formation in both intact and permeabilized cells, suggesting that the site of protein kinase C-mediated phosphorylation may be blocked in ET-1-stimulated Rat-1 cells. The results indicate that the receptor-G-protein-phospholipase C interaction for the two agonists may not conform to the same model.",
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Differences in the regulation of endothelin-1- and lysophosphatidic-acid-stimulated Ins(1,4,5)P3 formation in rat-1 fibroblasts. / Plevin, R; MacNulty, E E; Palmer, S; Wakelam, M J.

In: Biochemical journal, Vol. 280 , No. 3, 15.12.1991, p. 609-615.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Differences in the regulation of endothelin-1- and lysophosphatidic-acid-stimulated Ins(1,4,5)P3 formation in rat-1 fibroblasts

AU - Plevin, R

AU - MacNulty, E E

AU - Palmer, S

AU - Wakelam, M J

PY - 1991/12/15

Y1 - 1991/12/15

N2 - Endothelin-1 (ET-1)- and lysophosphatidic acid (LPA)-stimulated PtdIns(4,5)P2 hydrolysis has been studied in Rat-1 fibroblasts. Although both agonists caused the dose-dependent accumulation of inositol phosphates, a number of differences were observed. LPA induced a transient increase in Ins(1,4,5)P3 mass which returned to basal levels within 90 s, whereas the response to ET-1 did not desensitize, with levels remaining at 3-4 times basal values for up to 15 min. Stimulated decreases in mass levels of PtdIns(4,5)P2 mirrored Ins(1,4,5)P3 formation for both agonists. Experiments with electropermeabilized cells demonstrated that the effects of both agonists are stimulated by a phospholipase C controlled by a guanine-nucleotide-binding regulatory protein; however, there are differences in the nature of these interactions. The inositol phosphate response to ET-1 is poorly potentiated by guanosine 5'-[gamma-thio]triphosphate (GTP[S]) and markedly inhibited by guanosine 5'-[beta-thio]diphosphate (GDP[S]), whereas that to LPA is potentiated by GTP[S] but is relatively insensitive to GDP[S]. In addition, LPA decreased the lag time for the onset of GTP[S]-stimulated [3H]InsP3 accumulation, whereas ET-1 was without effect. Phorbol 12-myristate 13-acetate treatment of the cells inhibited LPA-stimulated, but not ET-1-stimulated, inositol phosphate formation in both intact and permeabilized cells, suggesting that the site of protein kinase C-mediated phosphorylation may be blocked in ET-1-stimulated Rat-1 cells. The results indicate that the receptor-G-protein-phospholipase C interaction for the two agonists may not conform to the same model.

AB - Endothelin-1 (ET-1)- and lysophosphatidic acid (LPA)-stimulated PtdIns(4,5)P2 hydrolysis has been studied in Rat-1 fibroblasts. Although both agonists caused the dose-dependent accumulation of inositol phosphates, a number of differences were observed. LPA induced a transient increase in Ins(1,4,5)P3 mass which returned to basal levels within 90 s, whereas the response to ET-1 did not desensitize, with levels remaining at 3-4 times basal values for up to 15 min. Stimulated decreases in mass levels of PtdIns(4,5)P2 mirrored Ins(1,4,5)P3 formation for both agonists. Experiments with electropermeabilized cells demonstrated that the effects of both agonists are stimulated by a phospholipase C controlled by a guanine-nucleotide-binding regulatory protein; however, there are differences in the nature of these interactions. The inositol phosphate response to ET-1 is poorly potentiated by guanosine 5'-[gamma-thio]triphosphate (GTP[S]) and markedly inhibited by guanosine 5'-[beta-thio]diphosphate (GDP[S]), whereas that to LPA is potentiated by GTP[S] but is relatively insensitive to GDP[S]. In addition, LPA decreased the lag time for the onset of GTP[S]-stimulated [3H]InsP3 accumulation, whereas ET-1 was without effect. Phorbol 12-myristate 13-acetate treatment of the cells inhibited LPA-stimulated, but not ET-1-stimulated, inositol phosphate formation in both intact and permeabilized cells, suggesting that the site of protein kinase C-mediated phosphorylation may be blocked in ET-1-stimulated Rat-1 cells. The results indicate that the receptor-G-protein-phospholipase C interaction for the two agonists may not conform to the same model.

KW - animals

KW - cell membrane permeability

KW - cells, cultured

KW - endothelins

KW - fibroblasts

KW - GTP-binding proteins

KW - guanosine 5'-O-(3-Thiotriphosphate)

KW - inositol phosphates

KW - lysophospholipids

KW - phosphatidylinositols

KW - phosphorylation

KW - rats

KW - tetradecanoylphorbol acetate

KW - type C phospholipases

UR - http://www.biochemj.org/bj/280/bj2800609.htm

M3 - Article

VL - 280

SP - 609

EP - 615

JO - Biochemical Journal

T2 - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 3

ER -