Protein Phosphatase-1 and Protein Phosphatase-2A from Rabbit Skeletal Muscle

Philip Cohen, Susana Alemany, Brian A. Therese, Therese J. Resink, Peter Stralfors, H. Y. Lim Tung

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    Abstract

    This chapter describes the purification of two forms of protein phosphatase- 1 (PP-I1 and PP-lG), and three forms of protein phosphatase-2A (PP-2A0, PP-2A1, and PP-2A2) from skeletal muscle. Procedures for isolating the free catalytic subunits (termed PP-lc and PP-2Ac) are documented and the structures of these enzymes are summarized in the chapter. The free catalytic subunits are isolated by a modification of the procedure of Lee and co-workers. Skeletal muscle extracts prepared from 3000 g of muscle are adjusted to pH 7.2 with 10 M ammonium hydroxide, and 350 g of solid ammonium sulfate are added to each liter of solution to bring the degree of saturation to 55%. After standing for 30 min the suspension is centrifuged for 40 min at 4200 g and the supernatant discarded. The PP-2Ac from poly(L-lysine)-Sepharose is purified in an identical manner to PP-Ic.

    Original languageEnglish
    Pages (from-to)390-408
    Number of pages19
    JournalMethods in Enzymology
    Volume159
    Issue numberC
    DOIs
    Publication statusPublished - 1988

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    Cohen, P., Alemany, S., Therese, B. A., Resink, T. J., Stralfors, P., & Lim Tung, H. Y. (1988). Protein Phosphatase-1 and Protein Phosphatase-2A from Rabbit Skeletal Muscle. Methods in Enzymology, 159(C), 390-408. https://doi.org/10.1016/0076-6879(88)59039-0