Asano T, Morishita R, Ohashi K, Nagahama M, Miyake T, Kato K
Institute for Developmental Research, Aichi Prefectural Colony, Japan.
J Neurochem. 1995 Mar;64(3):1267-73. doi: 10.1046/j.1471-4159.1995.64031267.x.
For a study of the localization of various forms of the gamma subunit of G proteins, antibodies were raised in rabbits against peptides that corresponded to partial amino acid sequences of bovine gamma 2, gamma 3, gamma 5, and gamma 7. Affinity-purified antibodies against gamma 2, gamma 3, and gamma 5 reacted specifically with gamma 2, gamma 3, and gamma 5, respectively, but the antibody against gamma 7 reacted with gamma 2, gamma 3, and a novel gamma subunit, designated gamma S1, as well as with gamma 7. Because these antibodies reacted with the respective forms of the gamma subunit from rat brain, we investigated the localization of gamma subunits in the rat. gamma 2 and gamma 3 were abundant in all regions in the brain, whereas the concentration of gamma 5 and gamma 7 was relatively low with the single exception being a high concentration of gamma 7 in the striatum. The concentration of gamma 2 was consistently high during ontogenic development in the rat brain, whereas gamma 3 appeared about a week after birth and their concentrations then increased until a month after birth. In tissues other than the brain, gamma 3 was observed only in the pituitary gland, whereas gamma 2, gamma 5, and gamma 7 were found in a variety of tissues. In addition, most tissues contained relatively high concentrations of some other gamma subunit, which was detected with an antibody against a gamma 7-derived peptide and appeared to be gamma S1. Among cloned cells tested, gamma 3 was detected only in PC12 pheochromocytoma cells.(ABSTRACT TRUNCATED AT 250 WORDS)
为了研究G蛋白γ亚基各种形式的定位,用对应于牛γ2、γ3、γ5和γ7部分氨基酸序列的肽段在兔体内制备抗体。抗γ2、γ3和γ5的亲和纯化抗体分别与γ2、γ3和γ5特异性反应,但抗γ7的抗体与γ2、γ3、一种新的γ亚基(命名为γS1)以及γ7都发生反应。由于这些抗体与大鼠脑中相应形式的γ亚基发生反应,我们研究了大鼠体内γ亚基的定位。γ2和γ3在脑的所有区域都很丰富,而γ5和γ7的浓度相对较低,唯一的例外是纹状体中γ7浓度较高。在大鼠脑的个体发育过程中,γ2的浓度始终很高,而γ3在出生后约一周出现,然后其浓度一直增加到出生后一个月。在脑以外的组织中,仅在垂体中观察到γ3,而在多种组织中发现了γ2、γ5和γ7。此外,大多数组织含有相对较高浓度的其他一些γ亚基,用抗γ7衍生肽的抗体检测到,似乎是γS1。在所测试的克隆细胞中,仅在PC12嗜铬细胞瘤细胞中检测到γ3。(摘要截短于250字)