Ca2+ channel activation by platelet-derived growth factor-induced tyrosine phosphorylation and Ras guanine triphosphate-binding proteins in rat glomerular mesangial cells

Heping Ma, Hiroshi Matsunaga, Bing Li, Bernhard Schieffer, Mario B Marrero, Brian N. Ling

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

We investigated the signaling pathways mediating 1-pS Ca2+ channel activation by PDGF in cultured rat mesangial cells. In cell-attached patches, intrapipette PDGF-BB (PDGF B chain homodimer isoform) (50 ng/ml) dramatically stimulates channel activity (P < 0.003, n = 6). Tyrosine kinase inhibition (100 μM genistein or 10 (μM tryphostin 9) abolished PDGF-induced channel activation (P < 0.02, n = 6). In excised patches, the effect of tyrosine kinase inhibition could be reversed by 200 μM GTPγS (P < 0.02, n = 4). In contrast, 200 μM GDPβS inhibited PDGF-induced channel activity (P < 0.04, n = 6). Pertussis toxin (250 ng/ml) had no effect on PDGF-induced channel activity (P = 0.45, n = 6). When excised patches were exposed to anti-Ras antibody (5 μg/ ml), PDGF-induced channel activity was abolished (P < 0.002, n = 11). Western immunoblots revealed that PDGF-BB binding stimulates the formation of a membrane-bound complex consisting of growth factor receptor-binding protein 2, son of sevenless, and the PDGF-β receptor. Complex formation was abolished by genistein. In mesangial cells, the intrinsic tyrosine kinase activity of the PDGF-β receptor stimulates the formation of a membrane-bound growth factor receptor-binding protein 2/son of sevenless/PDGF-β receptor complex and activation of the pertussis toxin-insensitive GTP-binding protein, p21-Ras, which leads to the opening of 1-pS Ca2+ channels.

Original languageEnglish (US)
Pages (from-to)2332-2341
Number of pages10
JournalJournal of Clinical Investigation
Volume97
Issue number10
DOIs
StatePublished - May 15 1996

Fingerprint

Platelet-Derived Growth Factor Receptors
Mesangial Cells
Genistein
Platelet-Derived Growth Factor
Pertussis Toxin
Guanine
Protein-Tyrosine Kinases
Tyrosine
GRB2 Adaptor Protein
Carrier Proteins
Phosphorylation
Proto-Oncogene Proteins p21(ras)
Membranes
Growth Factor Receptors
GTP-Binding Proteins
Anti-Idiotypic Antibodies
Protein Isoforms
Western Blotting
triphosphoric acid
platelet-derived growth factor BB

Keywords

  • Ca channels
  • G-proteins
  • Growth factor receptors
  • Ras
  • Tyrosine kinase

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Ca2+ channel activation by platelet-derived growth factor-induced tyrosine phosphorylation and Ras guanine triphosphate-binding proteins in rat glomerular mesangial cells. / Ma, Heping; Matsunaga, Hiroshi; Li, Bing; Schieffer, Bernhard; Marrero, Mario B; Ling, Brian N.

In: Journal of Clinical Investigation, Vol. 97, No. 10, 15.05.1996, p. 2332-2341.

Research output: Contribution to journalArticle

Ma, Heping ; Matsunaga, Hiroshi ; Li, Bing ; Schieffer, Bernhard ; Marrero, Mario B ; Ling, Brian N. / Ca2+ channel activation by platelet-derived growth factor-induced tyrosine phosphorylation and Ras guanine triphosphate-binding proteins in rat glomerular mesangial cells. In: Journal of Clinical Investigation. 1996 ; Vol. 97, No. 10. pp. 2332-2341.
@article{b8b46ea999ae4b8792f0bd4680b64559,
title = "Ca2+ channel activation by platelet-derived growth factor-induced tyrosine phosphorylation and Ras guanine triphosphate-binding proteins in rat glomerular mesangial cells",
abstract = "We investigated the signaling pathways mediating 1-pS Ca2+ channel activation by PDGF in cultured rat mesangial cells. In cell-attached patches, intrapipette PDGF-BB (PDGF B chain homodimer isoform) (50 ng/ml) dramatically stimulates channel activity (P < 0.003, n = 6). Tyrosine kinase inhibition (100 μM genistein or 10 (μM tryphostin 9) abolished PDGF-induced channel activation (P < 0.02, n = 6). In excised patches, the effect of tyrosine kinase inhibition could be reversed by 200 μM GTPγS (P < 0.02, n = 4). In contrast, 200 μM GDPβS inhibited PDGF-induced channel activity (P < 0.04, n = 6). Pertussis toxin (250 ng/ml) had no effect on PDGF-induced channel activity (P = 0.45, n = 6). When excised patches were exposed to anti-Ras antibody (5 μg/ ml), PDGF-induced channel activity was abolished (P < 0.002, n = 11). Western immunoblots revealed that PDGF-BB binding stimulates the formation of a membrane-bound complex consisting of growth factor receptor-binding protein 2, son of sevenless, and the PDGF-β receptor. Complex formation was abolished by genistein. In mesangial cells, the intrinsic tyrosine kinase activity of the PDGF-β receptor stimulates the formation of a membrane-bound growth factor receptor-binding protein 2/son of sevenless/PDGF-β receptor complex and activation of the pertussis toxin-insensitive GTP-binding protein, p21-Ras, which leads to the opening of 1-pS Ca2+ channels.",
keywords = "Ca channels, G-proteins, Growth factor receptors, Ras, Tyrosine kinase",
author = "Heping Ma and Hiroshi Matsunaga and Bing Li and Bernhard Schieffer and Marrero, {Mario B} and Ling, {Brian N.}",
year = "1996",
month = "5",
day = "15",
doi = "10.1172/JCI118676",
language = "English (US)",
volume = "97",
pages = "2332--2341",
journal = "Journal of Clinical Investigation",
issn = "0021-9738",
publisher = "The American Society for Clinical Investigation",
number = "10",

}

TY - JOUR

T1 - Ca2+ channel activation by platelet-derived growth factor-induced tyrosine phosphorylation and Ras guanine triphosphate-binding proteins in rat glomerular mesangial cells

AU - Ma, Heping

AU - Matsunaga, Hiroshi

AU - Li, Bing

AU - Schieffer, Bernhard

AU - Marrero, Mario B

AU - Ling, Brian N.

PY - 1996/5/15

Y1 - 1996/5/15

N2 - We investigated the signaling pathways mediating 1-pS Ca2+ channel activation by PDGF in cultured rat mesangial cells. In cell-attached patches, intrapipette PDGF-BB (PDGF B chain homodimer isoform) (50 ng/ml) dramatically stimulates channel activity (P < 0.003, n = 6). Tyrosine kinase inhibition (100 μM genistein or 10 (μM tryphostin 9) abolished PDGF-induced channel activation (P < 0.02, n = 6). In excised patches, the effect of tyrosine kinase inhibition could be reversed by 200 μM GTPγS (P < 0.02, n = 4). In contrast, 200 μM GDPβS inhibited PDGF-induced channel activity (P < 0.04, n = 6). Pertussis toxin (250 ng/ml) had no effect on PDGF-induced channel activity (P = 0.45, n = 6). When excised patches were exposed to anti-Ras antibody (5 μg/ ml), PDGF-induced channel activity was abolished (P < 0.002, n = 11). Western immunoblots revealed that PDGF-BB binding stimulates the formation of a membrane-bound complex consisting of growth factor receptor-binding protein 2, son of sevenless, and the PDGF-β receptor. Complex formation was abolished by genistein. In mesangial cells, the intrinsic tyrosine kinase activity of the PDGF-β receptor stimulates the formation of a membrane-bound growth factor receptor-binding protein 2/son of sevenless/PDGF-β receptor complex and activation of the pertussis toxin-insensitive GTP-binding protein, p21-Ras, which leads to the opening of 1-pS Ca2+ channels.

AB - We investigated the signaling pathways mediating 1-pS Ca2+ channel activation by PDGF in cultured rat mesangial cells. In cell-attached patches, intrapipette PDGF-BB (PDGF B chain homodimer isoform) (50 ng/ml) dramatically stimulates channel activity (P < 0.003, n = 6). Tyrosine kinase inhibition (100 μM genistein or 10 (μM tryphostin 9) abolished PDGF-induced channel activation (P < 0.02, n = 6). In excised patches, the effect of tyrosine kinase inhibition could be reversed by 200 μM GTPγS (P < 0.02, n = 4). In contrast, 200 μM GDPβS inhibited PDGF-induced channel activity (P < 0.04, n = 6). Pertussis toxin (250 ng/ml) had no effect on PDGF-induced channel activity (P = 0.45, n = 6). When excised patches were exposed to anti-Ras antibody (5 μg/ ml), PDGF-induced channel activity was abolished (P < 0.002, n = 11). Western immunoblots revealed that PDGF-BB binding stimulates the formation of a membrane-bound complex consisting of growth factor receptor-binding protein 2, son of sevenless, and the PDGF-β receptor. Complex formation was abolished by genistein. In mesangial cells, the intrinsic tyrosine kinase activity of the PDGF-β receptor stimulates the formation of a membrane-bound growth factor receptor-binding protein 2/son of sevenless/PDGF-β receptor complex and activation of the pertussis toxin-insensitive GTP-binding protein, p21-Ras, which leads to the opening of 1-pS Ca2+ channels.

KW - Ca channels

KW - G-proteins

KW - Growth factor receptors

KW - Ras

KW - Tyrosine kinase

UR - http://www.scopus.com/inward/record.url?scp=0029664446&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029664446&partnerID=8YFLogxK

U2 - 10.1172/JCI118676

DO - 10.1172/JCI118676

M3 - Article

C2 - 8636414

AN - SCOPUS:0029664446

VL - 97

SP - 2332

EP - 2341

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

SN - 0021-9738

IS - 10

ER -