Activation of platelet-derived growth factor (PDGF) receptor-operated Ca2+ channel via tyrosine kinase and ras in mesangial cells

H. Ma, H. Matsunaga, B. Li, M. B. Marrero, D. C. Eaton, B. N. Ling

Research output: Contribution to journalArticle

Abstract

We investigated the regulation of 1 pS Ca2+ channels by PDGF in cultured rat mesangial cells. In cell-attached patches, intrapipette PDGF-BB (50 ng/ml) increased NPo (channel number X open probability) from 0.0002 ± 0.0001 to 0.33 ±0.02 (mean ±SE; P<0.003; n=6). Tyrosine kinase inhibition ( 100 uM genistein or 10 uM tryphostin9) abolished PDGF-induced channel activation (P<0.02; n=6). In inside-out patches, the effect of tyrosine kinase inhibition could be reversed by cytoplasmic exposure to a non-hydrolyzable GTP analogue (200 μM GTPyS) (P<0.02; n=4). In contrast, 200 μM GDPβS inhibited PDGF-induced channels (P<0.04; n=6). Pertussis toxin (250 ng/mi PTX) had no effect on PDGF-induced channel activity (P=0.45; n=6). When excised patches were exposed to anti-Aas antibody (5 ug/ml), PDGF-induced channel activity was abolished (PO.007; n=6). Western immunoblots revealed that genistein blocks PDGF-induced membrane translocation of growth factor receptor-binding protein 2 and the guanine nucleotide exchange factor, Son of Sevenless, to form a complex with the PDGFβ receptor itself. In mesangial cells, intrinsic tyrosine kinase activity of the PDGFβ receptor stimulates 1 pS Ca2+ channels via downstream activation of the PTX-insensitive GTP-binding protein, p21-Ras.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number3
StatePublished - Dec 1 1996

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Platelet-Derived Growth Factor Receptors
Mesangial Cells
Platelet-Derived Growth Factor
Protein-Tyrosine Kinases
Chemical activation
Genistein
Guanine Nucleotide Exchange Factors
Growth Factor Receptors
Pertussis Toxin
Guanosine Triphosphate
GTP-Binding Proteins
Rats
Anti-Idiotypic Antibodies
Carrier Proteins
Western Blotting
Membranes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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Activation of platelet-derived growth factor (PDGF) receptor-operated Ca2+ channel via tyrosine kinase and ras in mesangial cells. / Ma, H.; Matsunaga, H.; Li, B.; Marrero, M. B.; Eaton, D. C.; Ling, B. N.

In: FASEB Journal, Vol. 10, No. 3, 01.12.1996.

Research output: Contribution to journalArticle

Ma, H. ; Matsunaga, H. ; Li, B. ; Marrero, M. B. ; Eaton, D. C. ; Ling, B. N. / Activation of platelet-derived growth factor (PDGF) receptor-operated Ca2+ channel via tyrosine kinase and ras in mesangial cells. In: FASEB Journal. 1996 ; Vol. 10, No. 3.
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