Structure-function relationships of cation translocation by Ca(2+)- and Na+, K(+)-ATPases studied by site-directed mutagenesis.

Site-directed mutagenesis studies of the sarcoplasmic reticulum Ca(2+)-ATPase have pinpointed five amino acid residues that are essential to Ca2+ occlusion, and these residues have been assigned to different parts of a Ca2+ binding pocket with channel-like structure. Three of the homologous Na+, K(+)-ATPase residues have been shown to be important for binding of cytoplasmic Na+ at transport sites. In addition, three of the above mentioned Ca(2+)-ATPase residues appear to participate in the countertransport of H+, and two of the Na+, K(+)-ATPase residues to participate in the countertransport of K+. Residues involved in energy transducing conformational changes have also been identified by mutagenesis. In the Ca(2+)-ATPase, ATP hydrolysis is uncoupled from Ca2+ transport following mutation of a tyrosine residue located at the top of transmembrane segment M5. This tyrosine, present also in the Na+, K(+)-ATPase, may play a critical role in closing the gate to a transmembrane channel.