Conformation of Immunoglobulin M. III. Structural requirements of antigen for complement fixation by equine IgM.
Complexes of IgM equine anti-dansyl antibodies and different dansyl substituted carriers were tested for their ability to fix complement (C). Only dansyl92-Ficoll and dansyl12-poly-L-lysine were found to be effective. Dansyl13-bovine serum albumin, dansyl127-keyhole limpet hemocyanin, and reduced and alkylated dansyl10-ribonuclease were all ineffective. Lack of C fixation by the dansyl-ribonuclease was not due to lack of antibody-antigen complex formation, since binding at the concentrations employed for C fixation was established. However, in contrast, polymerized dansyl-ribonuclease (polydisperse, with m.w. = 74,000 to 230,000) was very effective in inducing C fixation. These results suggest that large antigen size is necessary for IgM to bind in a multivalent fashion to provide the correct conformation for C fixation. A similar conclusion had been made in earlier studies on rabbit IgM by Cunniff and Stollar. Since optimal C fixation occurred at lower antigen concentrations than maximal precipitation, it would appear that complexes in which several combining sites within a given IgM molecule may be bound to the same antigenic surface may be the most effective. The observation that the amount of C1q bound to antibody was the same in the presence and absence of antigen suggests that enhanced C fixation by antibody-antigen complexes is due to additional C component interactions such as C1r or C1s.
D000818:Animals; D000936:Antigen-Antibody Complex; D000941:Antigens; D001666:Binding Sites, Antibody; D003172:Complement C1 / Q000276:immunology; D003168:Complement Fixation Tests; D003619:Dansyl Compounds / Q000276:immunology / Q000494:pharmacology; D006736:Horses; D007075:Immunoglobulin M; D011108:Polymers; D011487:Protein Conformation; D012260:Ribonucleases / Q000494:pharmacology
D000936:Antigen-Antibody Complex; D000941:Antigens; D003172:Complement C1; D003619:Dansyl Compounds; D007075:Immunoglobulin M; D011108:Polymers; D012260:Ribonucleases