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G(o) 型G蛋白的两种形式:在大鼠各种组织和克隆细胞中的鉴定与分布

Two forms of G(o) type G proteins: identification and distribution in various rat tissues and cloned cells.

作者信息

Asano T, Morishita R, Kato K

机构信息

Department of Biochemistry, Institute for Developmental Research, Aichi Prefectural Colony, Japan.

出版信息

J Neurochem. 1992 Jun;58(6):2176-81. doi: 10.1111/j.1471-4159.1992.tb10961.x.

Abstract

A G(o) type G protein distinct from the major species of G(o) was recently isolated from bovine brain and designated G(o)*. The cDNAs encoding two forms of mammalian G(o) alpha were also isolated and designated GoA alpha and GoB alpha. To recognize two forms of G(o) type G proteins, we raised antibodies in rabbits against two peptides with sequences found only in the respective proteins of murine GoA alpha (SNTYEDAAAYIQTQF) and GoB alpha (TEAVAHIQGQYWSK). Purified anti-GoA alpha antibodies reacted with the major species of G(o) alpha purified from bovine and rat brain, whereas anti-GoB alpha antibodies reacted only with rat G(o)*alpha, but not with the major species of G(o) alpha or bovine G(o)*alpha. These results indicate that the major species of G(o) alpha is encoded by GoA alpha cDNA and G(o)*alpha is encoded by GoB alpha cDNA. Using these antibodies, the distribution of GoA and GoB was studied in various rat tissues and cloned cells. Both GoA and GoB were present in many tissues, but their distribution in peripheral tissues was distinct. GoA alpha seemed to associate mainly with neural tissues. On the other hand, relatively high concentrations of GoB alpha were present in the brain, pituitary gland, adipose tissue, lung, and testis. The concentrations of both GoA and GoB in the brain increased during ontogenic development, but the increase in GoB was observed at a later age. Both GoA and GoB were found in such cloned cells as PC12, NG108-15, C6, GA-1, G8, and 3T3-L1 cells. Treatment of PC12 cells with nerve growth factor caused the extension of neuron-like processes and the increase in the level of GoA, but not in the level of GoB.

摘要

最近从牛脑中分离出一种不同于主要类型G(o)的G(o)型G蛋白,并将其命名为G(o)*。还分离出了编码两种形式的哺乳动物G(o)α的cDNA,并分别命名为GoAα和GoBα。为了识别两种形式的G(o)型G蛋白,我们用兔抗两种肽段的抗体进行免疫,这两种肽段的序列仅存在于鼠GoAα(SNTYEDAAAYIQTQF)和GoBα(TEAVAHIQGQYWSK)各自的蛋白中。纯化的抗GoAα抗体与从牛脑和大鼠脑中纯化的主要类型的G(o)α发生反应,而抗GoBα抗体仅与大鼠G(o)*α发生反应,与主要类型的G(o)α或牛G(o)*α均不发生反应。这些结果表明,主要类型的G(o)α由GoAα cDNA编码,而G(o)*α由GoBα cDNA编码。利用这些抗体,研究了GoA和GoB在各种大鼠组织和克隆细胞中的分布。GoA和GoB在许多组织中都有表达,但它们在外周组织中的分布不同。GoAα似乎主要与神经组织相关。另一方面,在脑、垂体、脂肪组织、肺和睾丸中存在相对较高浓度的GoBα。在个体发育过程中,脑中GoA和GoB的浓度均升高,但GoB的升高出现在较晚的年龄。在PC12、NG108-15、C6、GA-1、G8和3T3-L1等克隆细胞中均发现了GoA和GoB。用神经生长因子处理PC12细胞会导致神经元样突起的延伸和GoA水平的升高,但不会导致GoB水平的升高。

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