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G蛋白αi或αq亚基表达或激活的稳定变化会影响β1和β2亚基的表达。

Stable changes in expression or activation of G protein alpha i or alpha q subunits affect the expression of both beta 1 and beta 2 subunits.

作者信息

Hermouet S, Murakami T, Spiegel A M

机构信息

Molecular Pathophysiology Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.

出版信息

FEBS Lett. 1993 Jul 26;327(2):183-8. doi: 10.1016/0014-5793(93)80166-r.

Abstract

G proteins consist of three subunits: alpha, beta and gamma. Four beta subunits have been cloned: beta 1 and beta 4 (36 kDa), and beta 2 and beta 3 (35 kDa). We studied endogenous beta subunits in mouse NIH 3T3 fibroblasts stably expressing high levels of G protein alpha subunits after transfection with cDNAs encoding alpha i1, alpha i2, alpha i3 and alpha q. Immunoblots showed that NIH 3T3 cells express beta 36 and beta 35 subunits; in these cells, beta 35 subunits are four times more abundant than beta 36 subunits. We could detect beta 1 and beta 2 mRNA, but neither beta 3 nor beta 4 mRNA. We found that a stable increase in expression of wild-type alpha i1, alpha i2, alpha i3 or alpha q subunits is always accompanied by an increase in beta 1 and beta 2 mRNA and protein levels. There was no evidence of selectivity for an increase in beta 1 rather than beta 2 subunits depending on the type of alpha subunit overexpressed. However, constitutive activation or inactivation of alpha subunits induced specific changes in beta subunits. Expression of constitutively inactivated alpha i2 subunits was accompanied by an increase in mRNA and protein levels of both beta subunits. In contrast, cells expressing constitutively activated alpha i2 subunits did not show any change in the amount of beta proteins expressed in membranes, despite a significant increase in beta 1 and beta 2 mRNA. We conclude that stable changes in the levels of expression or degree of activation of G alpha subunits affect the level of expression, and possibly the turn-over, of beta subunits, without selectivity among beta 1 and beta 2 subunits.

摘要

G蛋白由三个亚基组成:α、β和γ。已克隆出四种β亚基:β1和β4(36 kDa),以及β2和β3(35 kDa)。我们研究了在稳定表达高水平G蛋白α亚基的小鼠NIH 3T3成纤维细胞中的内源性β亚基,这些细胞在用编码αi1、αi2、αi3和αq的cDNA转染后。免疫印迹显示NIH 3T3细胞表达β36和β35亚基;在这些细胞中,β35亚基的丰度是β36亚基的四倍。我们能够检测到β1和β2的mRNA,但检测不到β3和β4的mRNA。我们发现野生型αi1、αi2、αi3或αq亚基表达的稳定增加总是伴随着β1和β2的mRNA及蛋白水平的增加。没有证据表明根据过表达的α亚基类型,β1亚基的增加而非β2亚基的增加具有选择性。然而,α亚基的组成型激活或失活会诱导β亚基的特异性变化。组成型失活的αi2亚基的表达伴随着两种β亚基的mRNA和蛋白水平的增加。相反,表达组成型激活的αi2亚基的细胞,尽管β1和β2的mRNA显著增加,但膜中表达的β蛋白量没有任何变化。我们得出结论,Gα亚基表达水平或激活程度的稳定变化会影响β亚基的表达水平,可能还会影响其周转,且在β1和β2亚基之间没有选择性。

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