将注射到非洲爪蟾卵母细胞中的信使核糖核酸所表达的大鼠脑钠钾-ATP酶活性α-3亚基

Expression of active alpha-3 subunit of rat brain Na+,K+-ATPase from the messenger RNA injected into Xenopus oocytes.

作者信息

Hara Y, Ohtsubo M, Kojima T, Noguchi S, Nakao M, Kawamura M

机构信息

Dept. of Biochemistry, Tokyo Medical and Dental University School of Medicine, Japan.

出版信息

Biochem Biophys Res Commun. 1989 Aug 30;163(1):102-5. doi: 10.1016/0006-291x(89)92104-9.

DOI:10.1016/0006-291x(89)92104-9

PMID:2549984

Abstract

Cloned cDNA encoding the so-far uncharacterized alpha-3 subunit of rat brain Na+,K+-ATPase (Hara et al. (1987) J. Biochem. 102, 43-58, Shull et al. (1986) Biochemistry 25, 8125-8132) was incorporated into a vector carrying the SP6 promoter. The mRNA produced in vitro was injected into Xenopus oocytes with the mRNA encoding the Na+,K+-ATPase beta subunit of Torpedo electroplax. Increased Na+,K+-ATPase activity in the oocyte membrane was observed. This newly expressed activity was inhibited by ouabain (Ki = 1.5 x 10(-7) M), suggesting that the alpha-3 subunit of rat brain Na+,K+-ATPase is a highly ouabain-sensitive catalytic subunit.

摘要

编码大鼠脑钠钾ATP酶迄今未被鉴定的α-3亚基的克隆互补DNA（Hara等人，（1987年）《生物化学杂志》102卷，43 - 58页；Shull等人，（1986年）《生物化学》25卷，8125 - 8132页）被插入到携带SP6启动子的载体中。体外产生的信使核糖核酸与编码电鳐电器官钠钾ATP酶β亚基的信使核糖核酸一起注射到非洲爪蟾卵母细胞中。观察到卵母细胞膜中钠钾ATP酶活性增加。这种新表达的活性被哇巴因抑制（抑制常数Ki = 1.5×10⁻⁷M），表明大鼠脑钠钾ATP酶的α-3亚基是一种对哇巴因高度敏感的催化亚基。