Insulin receptor and altered glucose transport in a monensin-resistant mutant of Chinese hamster ovary cell

✍ Scribed by Tadashi Seguchi; Akihiko Yoshimura; Mayumi Ono; Satoshi Shite; Michihiro Kasahara; Yosuke Ebina; William J. Rutter; Michihiko Kuwano

• Publisher: John Wiley and Sons
• Year: 1989
• Tongue: English
• Weight: 802 KB
• Volume: 139
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1041390203

✦ Synopsis

A monensin-resistant mutant Monr-31, derived from Chinese hamster ovary (CHO) cell line, has been shown to have a reduced number of insulin receptors and a reduction in glucose uptake in response to insulin. We have further investigated the possibility that altered glucose uptake in Monr-31 cells i s related to an alteration in the activity of the insulin receptor. Uptake of glucosamine, 2-deoxy-~-glucose, and 3-O-methyl-~-glucose in Monr-31 cells was one-half to one-third that of CHO cells. The cellular content of the glucose transporter in Monr-31 was reduced to about one-third that of C H O as assayed by use of an antiglucose transporter antibody. After transfection with the human insulin receptor cDNA, we obtained clones CIR-0 from CHO, and MIR-2 and MIR-15 from Monr-31; CIR-0 expressed a tenfold higher level of the insulin-binding activity than did CHO, and MIR-2 and MIR-15 expressed a 20-foold higher level than did Mon'-31. Glucose uptake in both CHO and CIR-0 was significantly enhanced by exogenous insulin, but not in Monr-31, MIR-2, and MIR-15. The P-subunits of insulin receptor in CHO, CIR-0, Monr-31, and MIR-2 were similarly phosphorylated. The decreased glucose transport activity in Monr-3 1 cells is discussed in relation to the absence or presence of insulin receptor expression.