Complement-mediated hemolytic activity of succinylated concanavalin A: preparation and activity of cell intermediates.
Succinylated concanavalin A (SCon A) lyses sheep erythrocytes (E) in the presence of complement, whereas the native tetravalent lectin, Con A, is inactive. We have studied the ability of E-SCon A (ES) to interact with early acting guinea pig (gp) or human (hu) complement components (C1, C2, C4) and found that cell intermediates ESC1, ESC4, ESC14, and ESC142 can be generated that are analogous to intermediates conventionally prepared with E and rabbit IgM (pentameric) anti-Forssman antibody. Titration of gp or hu C1, C4, and C2, and quantification of the number of activated C1 molecules bound to ESC4 by the C1 fixation and transfer test showed in each case that an average of one effective site per cell was sufficient to cause cell lysis. Determination of tmax for optimal formation of ESC142 sites depended on the species combination of components used to make the intermediates, and the decay of ESC142 and EAC142 sites or sites generated with ESC4, EAC4, and trypsin-activated C2 were similar. The sugar alpha-D-methylglucopyranoside (alpha-MGP) inhibited binding of SCon A to E and eluted the lectin from ES, whereas galactose was nearly inactive, consistent with lectin sugar-binding selectivity. In contrast, both sugars were ineffective in eluting SCon A or C4hu from ESC4hu, indicating that C4hu blocked the interaction between lectin and alpha-MGP, perhaps by steric hindrance. SCon A is a divalent functional analogue of IgM anti-Forssman antibody that may be a uniquely suited reagent specific for cell membrane glycoconjugates for studying the mechanism of binding and activation of complement components.
D000818:Animals; D001666:Binding Sites, Antibody; D003167:Complement Activation; D003172:Complement C1 / Q000378:metabolism; D003175:Complement C2 / Q000378:metabolism; D003181:Complement C4 / Q000378:metabolism; D003168:Complement Fixation Tests; D003208:Concanavalin A / Q000031:analogs & derivatives* / Q000097:blood / Q000276:immunology / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism*; D006168:Guinea Pigs; D006461:Hemolysis; D006801:Humans; D066298:In Vitro Techniques; D008757:Methylglucosides / Q000378:metabolism; D051381:Rats
D003172:Complement C1; D003175:Complement C2; D003181:Complement C4; D008757:Methylglucosides; D003208:Concanavalin A; C027020:methylglucoside; C019558:succinylconcanavalin A