Contact regions for dinitrophenyl and menadione haptens in an immunoglobulin binding more than one antigen.
Protein 460 is a mouse myeloma gamma A(2) protein that competitively binds two small haptens, 2,4-epsilon-dinitrophenyl-L-lysine (Dnp-Lys) and 2-methyl-1:4-naphthaquinone thioglycollate (MenTG), to the antibody-combining region. The intact protein has a relatively inaccessible sulfhydryl group on each heavy chain. When it is substituted with a bulky reagent the binding affinity for MenTG decreases, while the binding of Dnp-Lys remains the same. Guanidine.HCl selectively reduces binding of Dnp-Lys; dimethylsulfoxide selectively reduces binding of MenTG. Papain digestion of protein 460 followed by column chromatography gave two fractions: one contained both binding activities and the other contained the sulfhydryl group. The affinity for Dnp-Lys of the first fraction is the same as that of the whole molecule, while affinity for MenTG is decreased. Since selective alteration of one or the other binding activity can occur in different ways, it seems likely that even though the haptens compete with each other, there is some spatial separation between the groups of contact amino-acid residues involved in the binding of these two haptens. These findings do not support the hypothesis that an immunoglobulin molecule carries combining sites complementary only to a single hapten or to a structurally related series of haptens, but rather suggests that the antibody-combining site may be a polyfunctional region capable of binding several structurally dissimilar haptens. We discuss a mechanism whereby polyfunctional combining sites can give rise to an antibody population (immune serum) that has a high degree of specificity to a single hapten.
D000478:Alkylation; D000818:Animals; D000918:Antibody Specificity; D000921:Antibody-Producing Cells / Q000378:metabolism; D001665:Binding Sites; D002460:Cell Line; D004140:Dinitrophenols / Q000378:metabolism*; D000939:Epitopes / Q000378:metabolism*; D006241:Haptens / Q000378:metabolism*; D007106:Immune Sera; D007136:Immunoglobulins / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D009194:Myeloma Proteins / Q000378:metabolism*; D011485:Protein Binding; D011489:Protein Denaturation; D011817:Rabbits; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013864:Thioglycolates / Q000378:metabolism; D014812:Vitamin K / Q000378:metabolism*
D004140:Dinitrophenols; D000939:Epitopes; D006241:Haptens; D007106:Immune Sera; D007136:Immunoglobulins; D009194:Myeloma Proteins; D013438:Sulfhydryl Compounds; D013864:Thioglycolates; D014812:Vitamin K; D008239:Lysine