Expression of the human erythrocyte glucose transporter in Escherichia coli.

Détails

ID Serval
serval:BIB_8E574D56C463
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Expression of the human erythrocyte glucose transporter in Escherichia coli.
Périodique
Proceedings of the National Academy of Sciences of the United States of America
Auteur(s)
Sarkar H.K., Thorens B., Lodish H.F., Kaback H.R.
ISSN
0027-8424
Statut éditorial
Publié
Date de publication
08/1988
Peer-reviewed
Oui
Volume
85
Numéro
15
Pages
5463-5467
Langue
anglais
Résumé
The gene encoding the human erythrocyte glucose transporter, cloned from HepG2 hepatoma cells, was expressed in Escherichia coli by introducing a prokaryote-type ribosome binding site, subcloning the gene into the T7 promoter/T7 polymerase expression system, and transforming a strain that is defective in glucose transport. Cells bearing plasmids with the transporter gene take up 2-deoxy-D-glucose and D-glucose, unlike cells bearing plasmids without the transporter gene. Moreover, 2-deoxy-D-glucose uptake is inhibited by unlabeled D-glucose, cytochalasin B, or mercuric chloride but not by L-glucose. The glucose transport protein is inserted into the membrane of E. coli, as evidenced by immunoblotting experiments with two site-directed polyclonal antibodies, one directed against the COOH terminus of the glucose transporter and the other directed against a synthetic peptide containing amino acid residues 225-238. As detected with both antibodies, the protein migrates with apparent molecular mass of 34 kDa in sodium dodecyl sulfate/12% polyacrylamide, a size similar to that of the unglycosylated glucose-transport protein synthesized in vitro.
Mots-clé
Base Sequence, Carcinoma, Hepatocellular, Cloning, Molecular, Cytochalasin B, Deoxyglucose, Electrophoresis, Polyacrylamide Gel, Erythrocytes, Escherichia coli, Gene Expression Regulation, Glucose, Humans, Immunoassay, Liver Neoplasms, Mercuric Chloride, Molecular Sequence Data, Monosaccharide Transport Proteins, Plasmids, Tumor Cells, Cultured
Pubmed
Web of science
Open Access
Oui
Création de la notice
24/01/2008 14:41
Dernière modification de la notice
08/05/2019 21:52
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