| Abstract: | Positive cooperativity demonstrated by Scatchard plot analysis of concanavalin A (con A) binding was found with native or glutaraldehyde-fixed erythrocytes. This suggests that factors other than membrane changes might be involved in the apparent increase in receptor binding affinity with increasing site occupancy. The elution pattern of 125I-con A chromatographed on Bio-Gel P-150 with decreasing concentration showed a drop in average molecular weight compatible with con A dissociation to dimers, protomers, and protomer fragments. Similarly, the per cent of 125I-con A specifically binding to Sephadex G-75 fell with decreasing concentration of con A applied. The inclusion of unlabeled carrier con A suppressed the dissociation of labeled con A in Bio-Gel P-150 and increased the per cent binding to Sephadex G-75. Both labeled and unlabeled con A were multibanded in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. As previously reported, the three major bands are consistent with intact protomer (approximately 25,000 daltons) and two fragments (approximately 13,000 and 10,000 daltons) with minor bands representing undissociated species. These observations indicate that there is a concentration-dependent association of Con A subunits which contribute to the observed positive cooperativity of con A binding to erythrocytes. |
