The Insulin Receptor of Embryonic Chicken Cartilage*

Abstract

Highly purified plasma membranes have been obtained from embryonic chicken cartilage by physical means rather than enzymatic digestion. Rapid and reversible binding of [l2!iI]iodoinsulin to these membranes is demonstrated. Similar to the insulin-binding properties of rat liver and adipocytes and human mononuclear cells, optimal specific binding of insulin to chondrocyte plasma membranes has a sharp pH optimum at 8.0, and maximal binding occurs at 2-4 C. Analysis of equilibrium binding reveals a curvilinear Scatchard plot, whose high affinity segment generates a maximum affinity of 1.0 x 109 M“, and a receptor concentration of 0.4 pmol⁄mg membrane protein. This affinity constant is similar to those generated for insulin binding to membranes prepared from embryonic chicken liver (2.5 × 109 M“'•'), rat liver (1.4 X 109 M“1), and mouse liver (0.6 x 109 M'), whereas the receptor concentration is less than that of embryonic chicken liver membranes (1.1 pmol⁄mg), which in turn was less than those of rat liver me mbranes (2.8 pmol⁄mg) and mouse liver membranes (3.5 pmol⁄mg). Kinetic studies show augmentation of insulin-receptor dissociation by excess insulin when initial receptor occupancy is low, suggesting that negative cooperativity cooperativity is present. There is little or no interaction of other hormones with the chondrocyte insulin receptor, with the exception f proinsulin and the insulin-like growth factors. Porcine proinsulin, bovine proinsulin, somatomedin C, and nonsuppressible insulin-like protein prevent [l2ftI]iodoinsulin binding to chondrocyte plasma membranes with dose-response curves which are parallel to that of unlabeled porcine insulin itself, but with molar potencies relative to porcine insulin of 15%, 9%, 2.5%, and 1.4%, respectively. Porcine insulin and proinsulin both prevent binding of [12r'I]iodosomatomedin C to chondrocyte plasma membranes but with molar potencies less than 1% that of unlabeled somatomedin C These observations are consistent with the presence of a specific independent insulin receptor in embryonic chicken cartilage which is similar in its characteristics to the insulin receptor in previously described tissues. Insulin has a weak interaction with the chondrocyte receptor for somatomedin C. Interaction with the somatomedin receptor may be the mechanism by which insulin exerts anabolic effects on cartilage when used in pharmacological amounts