THE GLUCOSE-TRANSPORTER OF ESCHERICHIA-COLI - PURIFICATION AND CHARACTERIZATION BY NI+-CHELATE AFFINITY-CHROMATOGRAPHY OF THE IIBCGLC SUBUNIT

被引：37

作者：

WAEBER, U ^{[1
]}

BUHR, A ^{[1
]}

SCHUNK, T ^{[1
]}

ERNI, B ^{[1
]}

机构：

[1] UNIV MARBURG,DEPT BIOL,W-3550 MARBURG,GERMANY

来源：

FEBS LETTERS | 1993年 / 324卷 / 01期

关键词：

PHOSPHOTRANSFERASE SYSTEM; GLUCOSE CARRIER; NI+-CHELATE CHROMATOGRAPHY; ENZYME-II; ESCHERICHIA-COLI;

D O I：

10.1016/0014-5793(93)81542-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The IIBC(Glc) transmembrane subunit of the glucose transporter of E. coli containing a carboxy-terminal affinity tag consisting of six adjacent histidines was purified by nickel chelate affinity chromatography. The protein was constitutively overexpressed from a high copy number plasmid. 1.5 mg of 95% pure protein was obtained from 5 g (wet weight) cells. 70% of the phosphotransferase activity present in cell membranes was recovered. Adsorption to the nickel resin allows delipidation as well as rapid detergent exchange. The procedure was used to demonstrate exchange of subunits in the IIBC(Glc) dimer and it helds promise for the investigation of other protein-protein interactions.

引用

页码：109 / 112

页数：4

共 20 条

[1]

BUHR A, 1993, IN PRESS J BIOL CHEM

[2] PHOSPHORYLATION OF D-GLUCOSE IN ESCHERICHIA-COLI MUTANTS DEFECTIVE IN GLUCOSEPHOSPHOTRANSFERASE, MANNOSEPHOSPHOTRANSFERASE, AND GLUCOKINASE [J].