Modulation of the activity of the transverse tubule Mg2+ATPase from frog skeletal muscle by a monoclonal antibody in vitro

Mario S. Rosemblatt, Gonzalo Pérez, Enrique Jaimovich

Research output: Contribution to journalArticlepeer-review

Abstract

We have established several hybridoma lines that produce monoclonal antibodies against transverse tubule (t-tubule) proteins from frog skeletal muscle. The specificity of these antibodies was characterized by ELISA and Western immunoblotting with purified t-tubule, sarcoplasmic reticulum and partially purified sarcolemmal membranes. One of the monoclonal antibodies (2/34.4) recognizes a band of 109 000 Da in immunoblots. When purified frog t-tubule vesicles were preincubated with this antibody we observed an increase in the rate of the Mg2+-ATPase enzyme (up to six fold) which was dependent on antibody concentration. Immunocytological experiments done on cryostat sections of frog muscle indicate that the antigen recognized by this antibody is localized mainly at the level of the t-tubules (I band) and to a lesser extent at the sarcolemma. These results indicate that monoclonal antibody 2/34.4 recognizes the t-tubule Mg2+-ATPase and modulates its activity. This antibody should be useful as a probe on studies designed to study the physiological function of the enzyme.

Original languageEnglish
Pages (from-to)99-107
Number of pages9
JournalMolecular and Cellular Biochemistry
Volume106
Issue number2
DOIs
Publication statusPublished - Aug 1991

Keywords

  • frog skeletal muscle
  • Mg-ATPase
  • monoclonal antibodies
  • transverse tubules

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Clinical Biochemistry
  • Cell Biology
  • Medicine(all)

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