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Identification and partial characterization of a latent ATP, Mg-dependent protein phosphatase in rabbit skeletal muscle cytosol

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Summary

Fractionation of rabbit skeletal muscle cytosol on Aminohexyl-Sepharose has resulted in the identification of a latent ATP, Mg-dependent protein phosphatase whose catalytic subunit is in the active conformation, but is inhibited by the presence of more than one modulator unit. The partially purified enzyme is converted to an inactive, kinase FA-dependent form upon incubation at 30°C unless modulator-specific polyclonal antibodies are added to the preparation. The immunoglobulins also relieve the inhibition which is responsible for the low basal phosphatase activity of the enzyme, and they counteract all of the heat-stable inhibitor activity present in the preparation. Addition of free catalytic subunit abolishes the inhibition of the latent enzyme in a dose-dependent way, but cannot prevent the inactivation process. The inactivated phosphatase and the original latent enzyme exhibit the same apparent M r in sucrose density-gradient centrifugation (70 000) and in gel filtration (110 000).

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Abbreviations

PMSF:

Phenylmethanesulphonyl Fluoride

TLCK:

L-l-chloro-3-(4-tosylamido)-7-amino2-heptanone-hydrochloride

TPCK:

L-l-chloro-3-(4-tosvlamido)-4-phenyl-2-butanone

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Authors and Affiliations

  1. Afdeling Biochemie, Faculteit Geneeskunde, Katholieke Universiteit to Leuven, B-3000, Leuven, Belgium

    Jackie R. Vandenheede, Sigrid Staquet & Wilfried Merlevede

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  1. Jackie R. Vandenheede
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  2. Sigrid Staquet
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  3. Wilfried Merlevede
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Vandenheede, J.R., Staquet, S. & Merlevede, W. Identification and partial characterization of a latent ATP, Mg-dependent protein phosphatase in rabbit skeletal muscle cytosol. Mol Cell Biochem 87, 31–39 (1989). https://doi.org/10.1007/BF00421080

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  • DOI: https://doi.org/10.1007/BF00421080

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