The plasma membrane in the control of the signaling function of calcium

Eukaryotic plasma membranes contain three Ca-transporting systems: a Ca channel, an ATPase, and an Na/Ca exchanger. The ATPase is high-affinity, low-capacity system, which continuously pumps Ca out of cells. The Na/Ca exchanger is a low-affinity, high-capacity system, which is particularly active in excitable cells. The exchanger probably functions in both the Ca efflux and influx directions. The Ca-ATPase is a single polypeptide of Mr 138 kD, which is activated by calmodulin or, in its absence, by acidic phospholipids, polyunsaturated fatty acids, and limited proteolytic treatments. Trypsin produces a number of fragments, some of which (Mr 90, 85, and 81 kD) function as ATPases and transport Ca across reconstituted bilayer membranes. Trypsin proteolysis in the presence of different effectors has permitted us to locate the calmodulin-interacting domain of the enzyme in a 9-kD peripheral sequence that consists of a 4-kD calmodulin-binding subdomain and a subdomain of Mr 5 kD, which is essential for the expression of calmodulin stimulation. The Na/Ca exchanger of plasma membranes has not yet been identified with certainty. On the basis of purification attempts using different approaches, probable Mr's of 82, 70, or 33 kD have been proposed. Antibodies raised against the 33-kD protein partially inhibit the exchange activity of heart sarcolemma vesicles. They interact with the 33-kD protein, but also, under nonreducing conditions, with proteins of Mr approximately 70 and approximately 140 kD. Under reducing conditions, the reactivity with the latter component disappears. It is suggested that the monomeric Mr of the exchanger is 33 kD, and that intermolecular disulfide bridges associate monomers into dimeric and tetrameric forms.