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

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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.

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Abbreviations

t-tubules:

transverse-tubules

mAb:

monoclonal antibody

SR:

sarcoplasmic reticulum

SL:

sarcolemma

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Authors and Affiliations

  1. Laboratorio de Inmunología Celular, INTA Universidad de Chile, Casilla 138, Santiago, Chile

    Mario S. Rosemblatt & Gonzalo Pérez

  2. Departamento de Fisiología y Biofísica, Facultad de Medicina, Universidad de Chile and Centro de Estudios Científicos de Santiago, Chile

    Enrique Jaimovich

Authors
  1. Mario S. Rosemblatt
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  2. Gonzalo Pérez
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  3. Enrique Jaimovich
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Rosemblatt, M.S., Pérez, G. & Jaimovich, E. Modulation of the activity of the transverse tubule Mg2+ATPase from frog skeletal muscle by a monoclonal antibody in vitro . Mol Cell Biochem 106, 99–107 (1991). https://doi.org/10.1007/BF00230175

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  • DOI: https://doi.org/10.1007/BF00230175

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