Identification of Chicken Liver Glucose Transporter

Abstract

A cDNA, Ch-GT2, encoding a glucose transporter was cloned by screening a chicken liver λgt11 cDNA library and by modified polymerase chain reaction (PCR) techniques. The encoded protein is highly homologous to rat and human GLUT 2. Within the predicted amino acid sequence, there are 12 putative transmembrane helices with a relatively large exofacial hydrophilic loop between transmembrane segments 1 and 2, which is characteristic of mammalian GLUT 2. Expression of Ch-GT2 gene in E. coli results in sixfold increase in the uptake of 2-deoxy-D-glucose uptake, and this increment can be reduced by incubation with phloretin. Thus, Ch-GT2 is a bona fide chicken liver glucose transporter. A deletion mutant generated by PCR was expressed in bacteria and shown to be functional, indicating that the N-terminal seven amino acid residues of Ch-GT2 is not essential for transport of glucose. From Northern blot analysis, Ch-GT2 is prominently expressed in chicken liver with a transcript of 4.0 kb, but not in chicken brain and heart, suggesting that distinct types of glucose transporters are present in various chicken tissues.