Allosteric regulation by calcium of rabbit polyclonal anti-cyclic GMP antibody.
Calcium increased the binding of rabbit polyclonal antibodies and cGMP by increasing antibody affinity without altering the number of binding sites (Bmax). Competitive binding studies revealed that calcium increased the affinity of antibody for cGMP derivatives similarly, suggesting that the effects of this cation were antigen-independent. Kinetic binding studies demonstrated that calcium increased affinity by decreasing the dissociation rate without altering the association rate of antigen and antibody. Studies of the dissociation of antigen-antibody complexes preformed in the absence of calcium suggested that this cation regulated antibody function allosterically. These data contrast with those obtained previously suggesting that calcium regulated the interaction of cAMP and antibodies by increasing Bmax without altering affinity by reaction coupling. Re-analysis of those data demonstrated that calcium increased the affinity without altering the number of binding sites of antibodies to cAMP, in close agreement with the present results. These data suggest that allosteric modulation of antibody function by calcium may be a general mechanism regulating the interaction of polyclonal antibodies with cyclic nucleotides.
Patrinellis
A C
AC
Department of Medicine, Thomas Jefferson University, Philadelphia, PA 19107.
Waldman
S A
SA
D000494:Allosteric Regulation; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000915:Antibody Affinity / Q000187:drug effects; D000937:Antigen-Antibody Reactions / Q000187:drug effects; D001667:Binding, Competitive; D002118:Calcium / Q000494:pharmacology*; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000276:immunology*; D004533:Egtazic Acid / Q000494:pharmacology; D011817:Rabbits; D014443:Tyrosine / Q000031:analogs & derivatives / Q000276:immunology
D000906:Antibodies; D014443:Tyrosine; D004533:Egtazic Acid; C014832:succinyl cyclic GMP-tyrosine methyl ester; D006152:Cyclic GMP; D002118:Calcium