The glucose transporter of Escherichia coli

FEBS Letters

Buhr, Andreas¹  Waeber, Urs¹  Erni, Bernhard¹  Schunk, Thomas² 

[1] Institute for Biochemistry, University of Bern, Freiestr. 3, CH-3012 Bern, Switzerland;Department of Biology, University of Marburg, D-3500 Marburg, Germany

关键词: Phosphotransferase system;    Glucose carrier;    Ni+ chelate chromatography;    Enzyme II;    E. coli;    IIBCGlc (ptsG);    transmembrane subunit of the glucose transporter (gene);    IIAGlc (crr) cytoplasmic subunit of the glucose transporter (gene);    HABMan (manX);    IICMan (manY) and IIDMan (manZ);    subunits of the mannose transporter (genes). IIABCGlcNAc (nagE);    transporter for N-acetyl glucosamine (gene);    NTA;    nitrilotriacetic acid;   

DOI  :  10.1016/0014-5793(93)81542-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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 ofsubunits in the IIBC^Glc dimer and it helds promise for the investigation of other protein-protein interactions.

【 授权许可】

Unknown