A 1H-NMR comparison of calmodulin activation by calcium and by cadmium

Our previous reports based on pharmacological and histochemical evidence suggest that calcium and cadmium can both activate calmodulin (CaM)-dependent functions. The study reported here was carried out to explain these observations in molecular terms, using 400 MHz 1H-NMR. Changes in the spectrum of bovine brain CaM induced by 0 to 4 molar equivalents of calcium and cadmium were practically the same. In particular, the chemical shifts and line shape of signals due to Tyr-138, Phe-65, Phe-89 and Tml-115 were similarly affected by either ion. In addition, the effects of N-(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide (W-7, a CaM antagonist) on the phenylalanine aromatic regions, methionine methyl regions and high-field methyl regions of the spectra of both calcium- and cadmium-saturated proteins were practically identical. The effect of W-7 on calcium- and cadmium-saturated CaM was reflected in changes in the signals of Ile-27, Phe-68, Phe-92, Ile-100 and Val-142, as well as Met-71, Met-72, Met-76, Phe-89 and Phe-141. The results show that cadmium binds to all calcium-binding sites of CaM, and induces conformational changes that are as extensive as those brought about by calcium. W-7 also inhibits CaM activation by calcium and cadmium. Combined with our previous toxicological evidence, these results suggest that cadmium binds indiscriminately to CaM and that subsequent activation or modulation of CaM-dependent functions is confused as a result. This may be a mechanism contributing to cadmium poisoning.