Effects of octyl beta-glucoside on insulin binding to solubilized membrane receptors.
Octyl beta-glucoside (1%), a dialyzable detergent, was used to solubilize the insulin receptor of the turkey erythrocyte membrane. Insulin binding capacity was stable for at least 1 week when the receptor was kept in 1% octyl beta-glucoside at 4 degrees C. The binding properties of the solubilized receptor were examined at detergent concentrations above (1%) and below (0.6%) the critical micelle concentration. A reduction in insulin binding occurred when the detergent concentration was raised above the critical micelle concentration, due to an apparent decrease in the number of binding sites. The specificity of the receptor for insulin analogues was preserved, and the relative affinity of the solubilized receptor, desoctapeptide insulin greater than proinsulin greater than porcine insulin, was similar in 0.6% and 1% detergent. Addition of divalent cations increased insulin binding to a similar extent at both detergent concentrations, but there was a slightly greater stimulation of binding in 0.6% detergent as compared to 1% detergent. The pH optimum for binding was not affected by changes in the detergent concentration. These results indicate that the insulin receptor can be successfully solubilized by octyl beta-glucoside and that the binding activity is quite stable. Therefore, octyl beta-glucoside may be a useful detergent for purification of this receptor. In addition, the data indicate that the binding properties of the insulin receptor can be affected by changes in the physical state of the octyl beta-glucoside.
D000818:Animals; D003902:Detergents / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D005960:Glucosides / Q000494:pharmacology*; D006027:Glycosides / Q000494:pharmacology*; D007328:Insulin / Q000097:blood*; D007700:Kinetics; D008823:Micelles; D011972:Receptor, Insulin / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D013501:Surface-Active Agents / Q000494:pharmacology*; D014422:Turkeys
D003902:Detergents; D005960:Glucosides; D006027:Glycosides; D007328:Insulin; D008823:Micelles; D013501:Surface-Active Agents; C018619:octyl-beta-D-glucoside; D011972:Receptor, Insulin