The concanavalin A receptor from human erythrocytes in lipid bilayer membranes. Interaction with concanavalin A and succinyl-concanavalin A.
The concanavalin A receptor from human erythrocyte membranes has been isolated by affinity chromatography using the mild, readily-dialyzable detergent dodecyltrimethylammonium bromide. The purified protein has been reincorporated into large unilamellar phospholipid vesicles using a detergent dialysis technique. The mean diameter of these vesicles increases as the lipid: protein ratio decreases. Binding of succinyl-concanavalin A to these vesicles was quantitated using 125I-labelled lectin in a filtration assay. The concanavalin A receptor in lipid bilayer vesicles provides specific high affinity binding sites for succinyl-concanavalin A with an association constant of 2.13 . 10(6) M-1. Scatchard plots indicate positive cooperativity of binding at very low lectin concentrations, a characteristic also seen in concanavalin A binding to intact human erythrocytes. The presence of bovine serum albumin has little effect on lectin binding and is not required for expression of cooperativity. Concanavalin A effectively competes with succinyl-concanavalin A for binding to the vesicles with an association constant of 4.83 . 10(6) M-1. Receptor-bearing vesicles are readily agglutinated by concanavalin A but not by its succinylated derivative. The kinetics of vesicle agglutination are biphasic, with an initial rapid phase followed by a pseudo-first order process. We suggest that studies on reassembled receptor proteins in lipid bilayers can provide valuable insight into receptor involvement in transmembrane signalling events and the factors involved in cell membrane behaviour and cell agglutination.
D000371:Agglutination; D000818:Animals; D002417:Cattle; D003208:Concanavalin A / Q000031:analogs & derivatives* / Q000378:metabolism*; D004912:Erythrocytes / Q000032:analysis*; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism*; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D011952:Receptors, Concanavalin A / Q000097:blood* / Q000378:metabolism; D013997:Time Factors
D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D011952:Receptors, Concanavalin A; D003208:Concanavalin A; C019558:succinylconcanavalin A