Calcium binding to calmodulin. Cooperativity of the calcium-binding sites.
The effects of Mg2+ ion, pH, and KCl concentration on Ca2+ binding to calmodulin were studied by using a Ca2+ ion-sensitive electrode. The Ca2+ ion affinity of calmodulin increased with increasing pH or decreasing KCl concentration. Cooperativity between the Ca2+-binding sites was observed, and increased with decreasing pH or increasing KCl concentration. Free Ca2+ ion concentration was decreased by adding MgCl2 ion at low Mg2+ concentration and increased at higher concentrations in the presence of small amounts of Ca2+ ion. The decrease of free Ca2+ ion concentration by Mg2+ ion strongly suggests cooperativity between the Ca2+-binding sites, and it is difficult to explain the decrease in terms of the ordered binding models previously proposed. These results can be explained by a simple model which has four equivalent binding sites that bind Ca2+ and Mg2+ competitively, and showing cooperativity when either Ca2+ or Mg2+ is bound. Mg2+ ion binding to calmodulin was measured in the presence or absence of Ca2+ to confirm the validity of this model, and no Mg2+-specific site was observed.
D000495:Allosteric Site / Q000187:drug effects; D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D004533:Egtazic Acid / Q000494:pharmacology; D004566:Electrodes; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D008433:Mathematics; D008956:Models, Chemical; D011189:Potassium Chloride / Q000494:pharmacology
D002147:Calmodulin; D004533:Egtazic Acid; D011189:Potassium Chloride; D008274:Magnesium; D002118:Calcium