Adenosine triphosphatases during maturation of cultured human skeletal muscle cells and in adult human muscle

Biochim Biophys Acta. 1992 Nov 23;1112(1):89-98. doi: 10.1016/0005-2736(92)90258-n.


Na+/K(+)-ATPase, Mg(2+)-ATPase and sarcoplasmic reticulum (SR) Ca(2+)-ATPase are examined in cultured human skeletal muscle cells of different maturation grade and in human skeletal muscle. Na+/K(+)-ATPase is investigated by measuring ouabain binding and the activities of Na+/K(+)-ATPase and K(+)-dependent 3-O-methylfluorescein phosphatase (3-O-MFPase). SR Ca(2+)-ATPase is examined by ELISA, Ca(2+)-dependent phosphorylation and its activities on ATP and 3-O-methylfluorescein phosphate. Na+/K(+)-ATPase and SR Ca(2+)-ATPase are localized by immunocytochemistry. The activities of Na+/K(+)-ATPase and SR Ca(2+)-ATPase show a good correlation with the other assayed parameters of these ion pumps. All ATPase parameters investigated increase with the maturation grade of the cultured muscle cells. The number of ouabain-binding sites and the activities of Na+/K(+)-ATPase and K(+)-dependent 3-O-MFPase are significantly higher in cultured muscle cells than in muscle. The Mg(2+)-ATPase activity, the content of SR Ca(2+)-ATPase and the activities of SR Ca(2+)-ATPase and Ca(2+)-dependent 3-O-MFPase remain significantly lower in cultured cells than in muscle. The ouabain-binding constant and the molecular activities of Na+/K(+)-ATPase and SR Ca(2+)-ATPase are equal in muscle and cultured cells. During ageing of human muscle the activity as well as the concentration of SR Ca(2+)-ATPase decrease. Thus the changes of the activities of the ATPases are caused by variations of the number of their molecules. Na+/K(+)-ATPase is localized in the periphery of fast- and slow-twitch muscle fibers and at the sarcomeric I-band. SR Ca(2+)-ATPase is predominantly confined to the I-band, whereas fast-twitch fibers are much more immunoreactive than slow-twitch fibers. The presence of cross-striation for Na+/K(+)-ATPase and SR Ca(2+)-ATPase in highly matured cultured muscle cells indicate the development and subcellular organization of a transverse tubular system and SR, respectively, which resembles the in vivo situation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ca(2+) Mg(2+)-ATPase / metabolism*
  • Calcium-Transporting ATPases / metabolism*
  • Cells, Cultured
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunohistochemistry
  • Muscle Development
  • Muscles / cytology
  • Muscles / enzymology*
  • Ouabain / metabolism
  • Phosphorylation
  • Sarcoplasmic Reticulum / enzymology
  • Sodium-Potassium-Exchanging ATPase / metabolism*


  • Ouabain
  • 3-O-methyl-fluorescein phosphatase
  • Ca(2+) Mg(2+)-ATPase
  • Calcium-Transporting ATPases
  • Sodium-Potassium-Exchanging ATPase