Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues

Abstract

Complementary DNA clones encoding a facilitative glucose transporter-like protein have been isolated from human small intestine and muscle cDNA libraries. This 509-amino acid protein has 65.3, 54.3, and 57.5% identity with the previously described human erythrocyte/HepG2, liver, and fetal muscle glucose transporter/transporter-like proteins, respectively. RNA blotting studies indicate that transcripts encoding this protein are very abundant in adult human skeletal muscle and subcutaneous fat. The adult skeletal muscle glucose transporter-like protein was expressed in vitro by cDNA-directed transcription and cell-free translation of the synthetic mRNA. The in vitro-synthesized protein reacted with a monoclonal antibody, 1F8, which recognizes the insulin-regulatable glucose transporter expressed in rat skeletal muscle, heart, and adipocytes. In contrast, in vitro-synthesized erythrocyte/HepG2 and fetal muscle glucose transporters did not react with 1F8. The high levels in adult skeletal muscle and subcutaneous fat of mRNA encoding the adult skeletal muscle glucose transporter and its specific reactivity with monoclonal antibody 1F8 suggest that this protein is the major insulin-regulatable glucose transporter expressed in skeletal muscle and other insulin-responsive tissues.