Ca2+- and Mg2+-dependent ATPase activities in the deoxycholate-treated rat heart sarcolemma

1. Isolated rat heart sarcolemma was treated with different concentrations of an ionic detergent, deoxycholate (DOC) and ATP hydrolysis in the presence of Ca2+ or Mg2+ was determined. 2. Both Ca2+-dependent ATPase and Mg2+-dependent ATPase activities were decreased in the DOC-treated membranes; however, the depression of Mg2+-dependent ATPase activity was greater than that of Ca2+-dependent ATPase. 3. The differential changes in Ca2+-dependent ATPase and Mg2+-dependent ATPase activities were apparent when incubations with DOC were carried out for different time intervals and at different temperatures. 4. In DOC-treated preparations, the Km value for Ca2+-dependent ATPase was decreased whereas that for Mg2+-dependent ATPase was increased. The half maximal velocities of the Ca2+-dependent ATPase and Mg2+-dependent ATPase enzyme reactions in the treated preparations were obtained at a DOC: membrane protein ratio of 3.0 and 0.6, respectively. 5. In the DOC-treated membranes exhibiting the half maximal velocities of enzyme reactions, the Ka value for Ca2+-dependent ATPase was drastically reduced but remained unchanged for Mg2+-dependent ATPase. 6. The DOC treatment was associated with a loss of protein as well phospholipids and resulted in changes in the ultrastructural integrity of the membrane. 7. Varying degrees of decreases in the activities of sarcolemmal adenylate cyclase. (Na+-K+)-ATPase, 5'-nucleotidase and calcium binding were seen upon DOC treatment. 8. The extent of reduction in Ca2+-dependent ATPase and Mg2+-dependent ATPase activities were also different when the membrane was treated with a non-ionic detergent, Lubrol PX. 9. These data suggest that Ca2+-dependent ATPase in heart sarcolemma is more resistant than Mg2+-dependent ATPase to detergent treatments and further indicate some differences in the properties of these enzymes.