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首页> 外文期刊>American Journal of Physiology >Involvement of G protein betagamma-subunits in diverse signaling induced by G(i/o)-coupled receptors: study using the Xenopus oocyte expression system.
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Involvement of G protein betagamma-subunits in diverse signaling induced by G(i/o)-coupled receptors: study using the Xenopus oocyte expression system.

机译：G蛋白betagamma亚基参与由G（i / o）耦合受体诱导的多种信号传导：使用非洲爪蟾卵母细胞表达系统的研究。

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We studied the functions of betagamma-subunits of G(i/o) protein using the Xenopus oocyte expression system. Isoproterenol (ISO) elicited cAMP production and slowly activating Cl(-) currents in oocytes expressing beta(2)-adrenoceptor and the protein kinase A-dependent Cl(-) channel encoded by the cystic fibrosis transmembrane conductance regulator (CFTR) gene. 5-Hydroxytryptamine (5-HT), [d-Ala(2), d-Leu(5)]-enkephalin (DADLE), and baclofen enhanced ISO-induced cAMP levels and CFTR currents in oocytes expressing beta(2)-adrenoceptor-CFTR and 5-HT(1A) receptor (5-HT(1A)R), delta-opioid receptor, or GABA(B) receptor, respectively. 5-HT also enhanced pituitary adenylate cyclase activating peptide (PACAP) 38-induced cAMP levels and CFTR currents in oocytes expressing PACAP receptor, CFTR and 5-HT(1A)R. The 5-HT-induced enhancement of G(s)-coupled receptor-mediated currents was abrogated by pretreatment with pertussis toxin (PTX) and coexpression of G transducin alpha (G(t)alpha). The 5-HT-induced enhancement was further augmented by coexpression of the Gbetagamma-activated form of adenylate cyclase (AC) type II but not AC type III. Thus betagamma-subunits of G(i/o) protein contribute to the enhancement of G(s)-coupled receptor-mediated responses. 5-HT and DADLE did not elicit any currents in oocytes expressing 5-HT(1A)R or delta-opioid receptor alone. They elicited Ca(2+)-activated Cl(-) currents in oocytes coexpressing these receptors with the Gbetagamma-activated form of phospholipase C (PLC)-beta2 but not with PLC-beta1. These currents were inhibited by pretreatment with PTX and coexpression of G(t)alpha, suggesting that betagamma-subunits of G(i/o) protein activate PLC-beta2 and then cause intracellular Ca(2+) mobilization. Our results indicate that betagamma-subunits of G(i/o) protein participate in diverse intracellular signals, enhancement of G(s)-coupled receptor-mediated responses, and intracellular Ca(2+) mobilization.
机译：我们使用非洲爪蟾卵母细胞表达系统研究了G（i / o）蛋白的betagamma亚基的功能。异丙肾上腺素（ISO）在表达β（2）-肾上腺素受体和由囊性纤维化跨膜电导调节剂（CFTR）基因编码的蛋白激酶A依赖的Cl（-）通道的卵母细胞中诱导cAMP产生并缓慢激活Cl（-）电流。 5-羟色胺（5-HT），[d-Ala（2），d-Leu（5）]-脑啡肽（DADLE）和巴氯芬增强表达β（2）-肾上腺素受体的卵母细胞中ISO诱导的cAMP水平和CFTR电流-CFTR和5-HT（1A）受体（5-HT（1A）R），δ阿片受体或GABA（B）受体。 5-HT还增强了表达PACAP受体，CFTR和5-HT（1A）R的卵母细胞中垂体腺苷酸环化酶激活肽（PACAP）38诱导的cAMP水平和CFTR电流。通过用百日咳毒素（PTX）预处理和G转导蛋白α（G（t）alpha）的共表达，可以消除5-HT诱导的G（s）偶联受体介导的电流增强。 5-HT诱导的增强作用通过II型腺苷酸环化酶（AC）的Gbetagamma活化形式而不是III型AC的共表达而进一步增强。因此，G（i / o）蛋白的betagamma亚基有助于增强G（s）耦合受体介导的反应。 5-HT和DADLE不会在单独表达5-HT（1A）R或δ-阿片受体的卵母细胞中引发任何电流。他们在卵母细胞中引起Ca（2+）激活的Cl（-）电流与磷脂酶C（PLC）-beta2的Gbetagamma激活形式共表达这些受体，但与PLC-beta1无关。这些电流被PTX预处理和G（t）alpha的共表达抑制，表明G（i / o）蛋白的betagamma亚基激活PLC-beta2，然后引起细胞内Ca（2+）动员。我们的结果表明，Betagamma亚基的G（i / o）蛋白参与各种细胞内信号，G（s）耦合受体介导的反应和细胞内Ca（2+）动员的增强。

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