Reciprocal regulation of Gsα by palmitate and the βγ subunit

Reciprocal regulation of Gsα by palmitate and the βγ subunit
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棕榈酸酯和 βγ 亚基对 Gsα 的相互调节

DOI:
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发表时间:
1996
期刊:
影响因子:
--
通讯作者:
H. Bourne
H. Bourne
中科院分区:
--
文献类型:
--
作者:
T. Iiri;P. Backlund;T. L. Jones;P. Wedegaertner;H. Bourne

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G s的激素激活, 腺苷酸环化酶,促进α s从 Gβγ,加速共价连接的棕榈酸酯从 Gα亚基,并触发α s的一部分从质膜释放到胞质溶胶中。阐明 在这三个事件中,我们评估了 在Sf 9细胞制备的重组α s上附着的棕榈酸酯的体外。与未棕榈酰化的蛋白质相比, (从Sf 9细胞的胞质溶胶获得,用棕榈酰酯酶处理, 或表达为缺乏棕榈酰化位点的突变蛋白), 棕榈酰化α s(来自Sf 9膜,50%棕榈酰化) 分配到TX-114中表明,疏水性更强,并且 结合βγ的亲和力高5倍。βγ保护GDP结合 α s,而不是α s · GTP[γS],来自 通过重组酯酶的脱棕榈酰化。我们得出结论,βγ 结合和棕榈酰化相互增强, 促进α s的膜附着和解离 βγ的α s ·GTP可能介导 受体诱导的α s去棕榈酰化和 在完整细胞中蛋白质转移到胞质溶胶中。
Hormonal activation of G s , the stimulatory regulator of adenylyl cyclase, promotes dissociation of α s from Gβγ, accelerates removal of covalently attached palmitate from the Gα subunit, and triggers release of a fraction of α s from the plasma membrane into the cytosol. To elucidate relations among these three events, we assessed biochemical effects in vitro of attached palmitate on recombinant α s prepared from Sf9 cells. In comparison to the unpalmitoylated protein (obtained from cytosol of Sf9 cells, treated with a palmitoyl esterase, or expressed as a mutant protein lacking the site for palmitoylation), palmitoylated α s (from Sf9 membranes, 50% palmitoylated) was more hydrophobic, as indicated by partitioning into TX-114, and bound βγ with 5-fold higher affinity. βγ protected GDP-bound α s , but not α s · GTP[γS], from depalmitoylation by a recombinant esterase. We conclude that βγ binding and palmitoylation reciprocally potentiate each other in promoting membrane attachment of α s and that dissociation of α s ·GTP from βγ is likely to mediate receptor-induced α s depalmitoylation and translocation of the protein to cytosol in intact cells.
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