Association of (Ca + Mg)-ATPase activity with ATP-dependent Ca uptake in vesicles prepared from human erythrocytes.
Ghost membranes prepared from human erythrocytes exhibit 2 distinct (Ca + Mg)-ATPase1 activities (Quist and Roufogalis, Arch Biochem Biophys 168:240, 1975). (Ca + Mg)-ATPase activity dependent on a water soluble protein fraction is selectively lost from ghost membranes during preparation of vesicles under low ionic strength, slightly alkaline conditions. In this study, the Ca2+ dependence of the remaining membrane bound (Ca + Mg)-ATPase activity and ATP-dependent Ca uptake in vesicles were compared. The Ca2+ activation curves for (Ca + Mg)-ATPase activity and Ca uptake into vesicles were parallel over a Ca2+ range of 0.3-330 micrometer, and both curves have 2 apparent KA values for Ca2+ of 0.45 and 100 micrometer. Addition of a concentrated soluble protein fraction containing predominantly spectrin to the vesicles increased (Ca + Mg)-ATPase activity over twofold but did not affect the rate of Ca uptake. These findings suggest that the (Ca + Mg)-ATPase activity remaining in vesicles after extraction of the water soluble proteins is associated with the Ca pump whereas (Ca + Mg)-ATPase activity dependent on the soluble protein fraction is associated with some other function.
D000251:Adenosine Triphosphatases / Q000097:blood*; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D001693:Biological Transport, Active / Q000187:drug effects; D001798:Blood Proteins / Q000378:metabolism; D002118:Calcium / Q000097:blood*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology*
D001798:Blood Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D008274:Magnesium; D002118:Calcium