Functional differences in insulin receptors in rat adipocyte and human placenta membranes.
Incubation of human placenta membranes with low concentrations (0.1-0.2 mM) of dithiothreitol (DTT) increased insulin binding approximately 1.4-fold, while 10 mM DTT completely inhibited insulin binding. In contrast, treatment of rat adipocyte membranes with 0.5-2.0 mM DTT increased tracer insulin binding 3- to 6-fold, while higher levels of DTT (10 mM) also fully inhibited insulin binding. Scatchard analysis of insulin binding revealed that DTT treatment of adipocyte membranes resulted in an increase in both the high and low affinity dissociation constants. Purification of adipocyte insulin receptors by wheat germ agglutinin-Sepharose chromatography, followed by insulin-agarose affinity chromatography, resulted in loss of DTT stimulation of insulin binding. Comparison of insulin receptors purified from rat adipocytes or human placenta membranes revealed no significant differences in the DTT sensitivities of insulin binding or protein kinase activities. These data suggest that the functional properties of the rat adipocyte insulin receptor are modified by its membrane environment compared to those of insulin receptors in placenta membranes or purified insulin receptors in detergent solution.
Swanson
M L
ML
Department of Physiology and Biophysics, University of Iowa, Iowa City 52242.
Dudley
D T
DT
Walker
P S
PS
Boyle
T R
TR
Pessin
J E
JE
D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005978:Glutathione / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives / Q000378:metabolism; D007335:Insulin-Like Growth Factor II / Q000378:metabolism; D008297:Male; D010920:Placenta / Q000187:drug effects / Q000378:metabolism*; D011247:Pregnancy; D051381:Rats; D011972:Receptor, Insulin / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*
D007328:Insulin; C008147:insulin, iodo-; D007335:Insulin-Like Growth Factor II; D011972:Receptor, Insulin; D005978:Glutathione; D004229:Dithiothreitol