The calcium pump of plasma membranes.

Abstract

The calcium pump of plasma membranes is an ATPase of the E1E2 type; that is, it forms a phosphoenzyme during the reaction cycle and is inhibited by vanadate. It differs from the Ca2+-transporting ATPase of sarcoplasmic reticulum in molecular mass, immunological properties and Ca2+/ATP stoichiometry. Its affinity for calcium, which is low in the absence of calmodulin (Km, 10-20 microM), is increased by the latter (to a Km of about 0.5 microM). The effect of calmodulin is mimicked by acidic phospholipids (including the phosphorylated products of phosphatidylinositol), long-chain polyunsaturated fatty acids, and controlled treatment with a number of proteases. The ATPase has been purified to homogeneity from a number of plasma membranes using calmodulin affinity chromatography. The purified enzyme (a single polypeptide of molecular mass 138 kDa) pumps calcium into reconstituted liposomes in exchange for protons. Controlled trypsin proteolysis has shown that about one-third of the enzyme mass can be removed without impairing calcium transport. It has also indicated that the ability to bind calmodulin and to respond to it resides in a 9 kDa sequence of the enzyme molecule. The sequence contains a 4 kDa domain that binds calmodulin, and a 5 kDa domain which is essential for the stimulation.