title: The hormonal regulation and interaction of concanavalin A and insulin binding in R3230AC mammary adenocarcinoma cells.
abstract: To investigate the effects of concanavalin A on insulin binding to R3230AC mammary carcinomas, initial experiments were performed to characterize binding of concanavalin A. Concanavalin A binding was found to be specific and saturable. Equilibrium binding experiments demonstrated that addition of low concentrations of concanavalin A enhanced the binding of [3H]concanavalin A, suggestive of positively cooperative interactions. Binding of concanavalin A was responsive to hormonal alterations; tumor cells from diabetic rats showed enhanced binding of concanavalin A and insulin compared to cells from intact rats and administration of insulin to diabetic rats returned concanavalin A and insulin binding to levels seen in controls. Incubation of tumor cells with concanavalin A prior to addition of 125I-labelled insulin resulted in a reduction of insulin-binding capacity; succinyl-concanavalin A did not affect binding of insulin. The present inhibition of insulin binding by concanavalin A was highest at the lower insulin concentrations, providing a linearized Scatchard plot that yielded a calculated Kd value comparable to the low-affinity portion of the curvilinear Scatchard plot for insulin binding. The dissociation rate of bound insulin depended on receptor occupancy. Addition of concanavalin A after insulin binding reached equilibrium resulted in increased insulin binding at higher hormone concentrations, decreased rates of dissociation of insulin and a loss of the correlation between receptor occupancy and dissociation rates. Concanavalin A alone demonstrated an insulin-like effect on glucose transport, which in these tumor cells represents a decrease in transport of 3-O-methylglucose. These results suggest that binding of both concanavalin A and insulin to cells from this hormonally responsive neoplasm is under insulin regulation and demonstrates similar characteristics to those reported for a variety of normal cells. Furthermore, the interaction between concanavalin A and te cell membranes affects the affinity of the insulin receptor for insulin and appears to decrease the observed negative cooperativity.
mesh_terms: D019325:3-O-Methylglucose; D000230:Adenocarcinoma / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D003208:Concanavalin A / Q000378:metabolism* / Q000494:pharmacology; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism; D005260:Female; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism*; D008757:Methylglucosides / Q000378:metabolism; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism
chemical_list: D007328:Insulin; D008757:Methylglucosides; D003208:Concanavalin A; D019325:3-O-Methylglucose; D011972:Receptor, Insulin