Fukumoto H, Kayano T, Buse J B, Edwards Y, Pilch P F, Bell G I, Seino S
Howard Hughes Medical Institute, Chicago, Illinois.
J Biol Chem. 1989 May 15;264(14):7776-9.
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.
已从人小肠和肌肉cDNA文库中分离出编码一种易化性葡萄糖转运蛋白样蛋白的互补DNA克隆。这种由509个氨基酸组成的蛋白,与先前描述的人红细胞/HepG2、肝脏和胎儿肌肉葡萄糖转运蛋白/转运蛋白样蛋白分别具有65.3%、54.3%和57.5%的同源性。RNA印迹研究表明,编码该蛋白的转录本在成人骨骼肌和皮下脂肪中非常丰富。通过cDNA指导的合成mRNA转录和无细胞翻译,在体外表达了成人骨骼肌葡萄糖转运蛋白样蛋白。体外合成的蛋白与单克隆抗体1F8发生反应,该抗体可识别在大鼠骨骼肌、心脏和脂肪细胞中表达的胰岛素可调节葡萄糖转运蛋白。相比之下,体外合成的红细胞/HepG2和胎儿肌肉葡萄糖转运蛋白不与1F8发生反应。编码成人骨骼肌葡萄糖转运蛋白的mRNA在成人骨骼肌和皮下脂肪中的高水平及其与单克隆抗体1F8的特异性反应表明,该蛋白是在骨骼肌和其他胰岛素反应性组织中表达的主要胰岛素可调节葡萄糖转运蛋白。