Isolation of frog heart sarcolemma possessing (Ca2+ + Mg2+)-ATPase and Ca2+ pump activities

Biochim Biophys Acta. 1981 Apr 22;643(1):55-62. doi: 10.1016/0005-2736(81)90218-2.

Abstract

The presence of a (Ca2+ + Mg2+)-ATPase and Ca2+ pump in frog heart sarcolemma was investigated. Frog cardiac sarcolemma was isolated by using a modification of a recent procedure for isolating sarcolemma from dog heart (Morcos, N.C. and Drummond, G.I. (1980) Biochim. Biophys. Acta 598, 17-39). Despite the sparseness of sarcoplasmic reticulum in frog heart, this tissue possessed (Ca2+ + Mg2+)-ATPase activity which appeared endogenous to the sarcolemma. The purified sarcolemmal vesicles also possessed ouabain-sensitive (Na+ + K+)-ATPase, Na+ -Ca2+ exchange activity and Ca2+ pump activity. Inclusion of ionophore A23187 into the assay medium resulted in an approximately 5-fold increase in (Ca2+ + Mg2+)-ATPase which may be due to removal of inhibitory concentrations of Ca2+ accumulated within the membrane vesicles by an ATP-dependent process. Vesicles pre-loaded with 45Ca2+, via Na+-Ca2+ exchange, lost 92% of their Ca2+ when ATP was present and only 60% in the absence of ATP. The results indicate that frog heart sarcolemma possesses a (Ca2+ + Mg2+)-ATPase and an ATP-dependent Ca2+ pump which may be involved in the outward pumping of Ca2+ from the heart cell.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Biological Transport, Active / drug effects
  • Ca(2+) Mg(2+)-ATPase
  • Calcimycin / pharmacology
  • Calcium / metabolism*
  • Calcium-Transporting ATPases / metabolism*
  • Kinetics
  • Myocardium / metabolism*
  • Ouabain / pharmacology
  • Rana catesbeiana
  • Sarcolemma / drug effects
  • Sarcolemma / metabolism*
  • Sodium-Potassium-Exchanging ATPase / metabolism

Substances

  • Calcimycin
  • Ouabain
  • Ca(2+) Mg(2+)-ATPase
  • Calcium-Transporting ATPases
  • Sodium-Potassium-Exchanging ATPase
  • Calcium