Insulin receptor and altered glucose transport in a monensin-resistant mutant of Chinese hamster ovary cell.
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 is related to an alteration in the activity of the insulin receptor. Uptake of glucosamine, 2-deoxy-D-glucose, and 3-O-methyl-D-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 CHO 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-fold higher level than did Monr-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 beta-subunits of insulin receptor in CHO, CIR-0, Monr-31, and MIR-2 were similarly phosphorylated. The decreased glucose transport activity in Monr-31 cells is discussed in relation to the absence or presence of insulin receptor expression.
Seguchi
T
T
Department of Biochemistry, Oita Medical School, Japan.
Yoshimura
A
A
Ono
M
M
Shite
S
S
Kasahara
M
M
Ebina
Y
Y
Rutter
W J
WJ
Kuwano
M
M
D019325:3-O-Methylglucose; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D003847:Deoxyglucose / Q000493:pharmacokinetics; D004351:Drug Resistance; D005947:Glucose / Q000493:pharmacokinetics*; D008565:Membrane Proteins / Q000378:metabolism; D008757:Methylglucosides / Q000493:pharmacokinetics; D008985:Monensin; D009154:Mutation; D011233:Precipitin Tests; D011972:Receptor, Insulin / Q000378:metabolism*; D014162:Transfection
D008565:Membrane Proteins; D008757:Methylglucosides; D019325:3-O-Methylglucose; D008985:Monensin; D003847:Deoxyglucose; D011972:Receptor, Insulin; D005947:Glucose