利用基因融合和胰蛋白酶敏感插入法分析F因子TraD膜拓扑结构。
Analysis of F factor TraD membrane topology by use of gene fusions and trypsin-sensitive insertions.
作者信息
Lee M H, Kosuk N, Bailey J, Traxler B, Manoil C
机构信息
Departments of Genetics, University of Washington, Seattle, Washington 98195-7360, USA.
出版信息
J Bacteriol. 1999 Oct;181(19):6108-13. doi: 10.1128/JB.181.19.6108-6113.1999.
Abstract
This report describes a procedure for characterizing membrane protein topology which combines the analysis of reporter protein hybrids and trypsin-sensitive 31-amino-acid insertions generated by using transposons ISphoA/in and ISlacZ/in. Studies of the F factor TraD protein imply that the protein takes on a structure with two membrane-spanning sequences and amino and carboxyl termini facing the cytoplasm. It was possible to assign the subcellular location of one region for which the behavior of fused reporter proteins was ambiguous, based on the trypsin cleavage behavior of a 31-residue insertion.
摘要
本报告描述了一种用于表征膜蛋白拓扑结构的方法，该方法结合了对报告蛋白杂交体的分析以及使用转座子ISphoA/in和ISlacZ/in产生的对胰蛋白酶敏感的31个氨基酸插入片段的分析。对F因子TraD蛋白的研究表明，该蛋白具有两个跨膜序列，其氨基和羧基末端面向细胞质的结构。基于一个31个残基插入片段的胰蛋白酶切割行为，有可能确定一个区域的亚细胞定位，而融合报告蛋白在该区域的行为是不明确的。
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