对加州电鳐Na⁺/K⁺-ATP酶α亚基的催化磷酸化位点天冬氨酸-376和假定的ATP结合位点赖氨酸-507进行定点诱变。
Site-directed mutagenesis of Asp-376, the catalytic phosphorylation site, and Lys-507, the putative ATP-binding site, of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase.
作者信息
Ohtsubo M, Noguchi S, Takeda K, Morohashi M, Kawamura M
机构信息
Department of Biology, University of Occupational and Environmental Health, Kitakyushu, Japan.
出版信息
Biochim Biophys Acta. 1990 Jan 29;1021(2):157-60. doi: 10.1016/0005-2736(90)90028-m.
Point mutations of Asp-376 of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase (the site of phosphorylation during the catalytic cycle) to Asn, Glu or Thr led to virtual abolishment of Na+/K(+)-ATPase activity and ouabain-binding capacity. Replacement of Lys-507 of the same subunit (the putative ATP-binding site) by Met resulted in decreases in Na+/K(+)-ATPase activity and ouabain-binding capacity. These results are in agreement with those reported for rabbit sarcoplasmic reticulum Ca2(+)-ATPase (Maruyama, K. and MacLennan, D.H. (1988) Proc. Natl. Acad. Sci. USA 85, 3314-3318).
加州电鳐Na⁺/K⁺-ATP酶α亚基的天冬氨酸-376（催化循环中的磷酸化位点）点突变为天冬酰胺、谷氨酸或苏氨酸，导致Na⁺/K⁺-ATP酶活性和哇巴因结合能力几乎完全丧失。同一亚基的赖氨酸-507（假定的ATP结合位点）被甲硫氨酸取代，导致Na⁺/K⁺-ATP酶活性和哇巴因结合能力降低。这些结果与报道的兔肌浆网Ca²⁺-ATP酶的结果一致（丸山，K.和麦克伦南，D.H.（1988年）美国国家科学院院刊85，3314 - 3318）。
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