TY - JOUR
T1 - The G protein βγ subunit transduces the muscarinic receptor signal for Ca2+ release in Xenopus oocytes
AU - Stehno-Bittel, Lisa
AU - Krapivinsky, Grigory
AU - Krapivinsky, Lyubov
AU - Perez-Terzic, Carmen
AU - Clapham, David E.
PY - 1995/12/15
Y1 - 1995/12/15
N2 - At least 30 G protein-linked receptors stimulate phosphatidylinositol 4,5-bisphosphate phosphodiesterase (phospholipase Cβ, PLCβ) through G protein subunits to release intracellular calcium from the endoplasmic reticulum (Clapham, D. E. (1995) Cell 80, 259-268). Although both Gα and Gβγ G protein subunits have been shown to activate purified PLCβ in vitro, Gαq has been presumed to mediate the pertussis toxin-insensitive response in vivo. In this study, we show that Gβγ plays a dominant role in muscarinic-mediated activation of PLCβ by employing the Xenopus oocyte expression system. Antisense nucleotides and antibodies to Gαq/11 blocked the m3-mediated signal transduction by inhibiting interaction of the muscarinic receptor with the G protein. Agents that specifically bound free Gβγ subunits (Gα-GDP and a β-adrenergic receptor kinase fragment) inhibited acetylcholine-induced signal transduction to PLCβ, and injection of Gβγ subunits into oocytes directly induced release of intracellular Ca2+. We conclude that receptor coupling specificity of the Gαq/Gβγ heterotrimer is determined by Gαq; Gβγ is the predominant signaling molecule activating oocyte PLCβ.
AB - At least 30 G protein-linked receptors stimulate phosphatidylinositol 4,5-bisphosphate phosphodiesterase (phospholipase Cβ, PLCβ) through G protein subunits to release intracellular calcium from the endoplasmic reticulum (Clapham, D. E. (1995) Cell 80, 259-268). Although both Gα and Gβγ G protein subunits have been shown to activate purified PLCβ in vitro, Gαq has been presumed to mediate the pertussis toxin-insensitive response in vivo. In this study, we show that Gβγ plays a dominant role in muscarinic-mediated activation of PLCβ by employing the Xenopus oocyte expression system. Antisense nucleotides and antibodies to Gαq/11 blocked the m3-mediated signal transduction by inhibiting interaction of the muscarinic receptor with the G protein. Agents that specifically bound free Gβγ subunits (Gα-GDP and a β-adrenergic receptor kinase fragment) inhibited acetylcholine-induced signal transduction to PLCβ, and injection of Gβγ subunits into oocytes directly induced release of intracellular Ca2+. We conclude that receptor coupling specificity of the Gαq/Gβγ heterotrimer is determined by Gαq; Gβγ is the predominant signaling molecule activating oocyte PLCβ.
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U2 - 10.1074/jbc.270.50.30068
DO - 10.1074/jbc.270.50.30068
M3 - Article
C2 - 8530411
AN - SCOPUS:0029610780
SN - 0021-9258
VL - 270
SP - 30068
EP - 30074
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 50
ER -