期刊文献

Functional expression of gastric H,K-ATPase using the baculovirus expression system 收藏

使用杆状病毒表达系统功能性表达胃H，K-ATP酶

原文求助发布源

作者

Corné H.W.Klaassen;Tom J.F.Van Uem;Mariëlle P.De Moel;Godelieve L.J.De Caluwé;Herman G.P.Swarts;Jan Joep H.H.M.De Pont;

作者单位

Department of Biochemistry, University of Nijmegen, Nijmegen, The Netherlands[*]Correspondence address: J.J.H.H.M. De Pont, Department of Biochemistry, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands. Fax: (31) (80) 540 525.

关键词

H,K-ATPase; Baculovirus expression system; Sf9 cell; In vitro expression; SCH 28080;

关键词译文

H，K-ATP酶;杆状病毒表达系统; Sf9细胞;体外表达; SCH 28080;

发布日期

30 August 1993

页码

277-282

DOI

10.1016/0014-5793(93)80237-O

来源信息

FEBS Letters ISSN：0014-5793, 1993年, 329卷, 3期, 277-282页

摘要

A novel approach to construct a single recombinant baculovirus expressing two protein subunits simultaneously by replacing polyhedrin as well as p10 coding sequences is described. The recombinant baculovirus expressed the α- as well as the β-subunit of the gastric H.K-ATPase. Sf9 cells infected with this virus exhibited a K⁺- and SCH 28080-sensitive ATP-dependent phosphorylation capacity in purified Sf9 membranes similar to Natlve H,K-ATPase. This activity was not present in control membranes containing only one of the two H,K-ATPase subunits. We therefore conclude that both subunits are essential for the phosphorylation capacity of H.K-ATPase.

摘要译文

描述了通过替换多角体蛋白以及p10编码序列来构建表达两个蛋白质亚基的单一重组杆状病毒的新方法。重组杆状病毒表达胃H.K-ATP酶的α-亚基和β-亚基。用该病毒感染的Sf9细胞在纯化的Sf9膜中表现出K sup + / sup - 和SCH28080-敏感的ATP依赖性磷酸化能力，类似于Natlve H，K-ATP酶。该活性不存在于仅含有两个H，K-ATP酶亚基中的一个的对照膜中。因此，我们得出结论，两个亚基对于H.K-ATP酶的磷酸化能力是必需的。

Corné H.W.Klaassen;Tom J.F.Van Uem;Mariëlle P.De Moel;Godelieve L.J.De Caluwé;Herman G.P.Swarts;Jan Joep H.H.M.De Pont;. Functional expression of gastric H,K-ATPase using the baculovirus expression system[J]. FEBS Letters, 1993,329(3): 277-282