利用转座子Tn10编码的金属四环素/H⁺反向转运蛋白的半胱氨酸扫描突变体确定跨膜区段

Determination of a transmembrane segment using cysteine-scanning mutants of transposon Tn10-encoded metal-tetracycline/H+ antiporter.

作者信息

Kimura T, Suzuki M, Sawai T, Yamaguchi A

机构信息

Department of Cell Membrane Biology, Osaka University, Japan.

出版信息

Biochemistry. 1996 Dec 10;35(49):15896-9. doi: 10.1021/bi961568g.

DOI:10.1021/bi961568g

PMID:8961955

Abstract

The metal-tetracycline/H+ antiporter is a bacterial plasma membrane protein belonging to the 12-transmembrane transporter family. The boundaries of membrane-embedded domains can be clearly determined by means of a method involving cysteine-scanning mutants and their reactivity to N-ethylmaleimide. The results for 37 cysteine-scanning mutants around putative transmembrane helix 9 indicate that the residues from 266 to 275 and from 296 to 298 form water extruding loops, while residues from 276 to 295 and from 299 to 302 are embedded in the membrane. This method is generally useful for determination of the transmembrane regions of a polytopic membrane protein.

摘要

金属-四环素/H⁺反向转运蛋白是一种属于12跨膜转运蛋白家族的细菌质膜蛋白。膜嵌入结构域的边界可以通过一种涉及半胱氨酸扫描突变体及其与N-乙基马来酰亚胺反应性的方法来明确确定。围绕假定跨膜螺旋9的37个半胱氨酸扫描突变体的结果表明，266至275位以及296至298位的残基形成了水挤出环，而276至295位以及299至302位的残基则嵌入膜中。该方法通常可用于确定多跨膜蛋白的跨膜区域。