Identification of a high-molecular-weight cadmium-binding protein in copper-resistant Bacillus acidocaldarius cells

Bacillus acidocaldarius grown in the presence of Cu++ was capable of accumulating the metal in the form of a protease-sensitive high molecular weight (HMW) moiety whose formation was inhibited by actinomycin D. Only cells preadapted in Cu++ were able to grow in a Cd(++)-containing medium. A cell-free extract from cadmium-stressed cells was fractionated by gel-permeation chromatography. The majority of cadmium was found associated with a HMW protein fraction which was further purified by anion exchange chromatography and high-performance liquid chromatography. The molecular weight of the purified protein was estimated to be 23,000 by SDS-PAGE. Amino acid analysis showed a low cysteine content and an abundance of aspartate and glutamate. It is likely that the cadmium-binding protein is an essential component of the mechanism mediating recovery from heavy metal toxicity.