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

doi:10.1016/0076-6879(88)59039-0

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-l_G), 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-l_c and PP-2A_c) 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-2A_c from poly(L-lysine)-Sepharose is purified in an identical manner to PP-I_c.

Original language	English
Pages (from-to)	390-408
Number of pages	19
Journal	Methods in Enzymology
Volume	159
Issue number	C
DOIs	https://doi.org/10.1016/0076-6879(88)59039-0
Publication status	Published - 1988

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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title = "Protein Phosphatase-1 and Protein Phosphatase-2A from Rabbit Skeletal Muscle",

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.",

author = "Philip Cohen and Susana Alemany and Therese, {Brian A.} and Resink, {Therese J.} and Peter Stralfors and {Lim Tung}, {H. Y.}",

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T1 - Protein Phosphatase-1 and Protein Phosphatase-2A from Rabbit Skeletal Muscle

AU - Cohen, Philip

AU - Alemany, Susana

AU - Therese, Brian A.

AU - Resink, Therese J.

AU - Stralfors, Peter

AU - Lim Tung, H. Y.

PY - 1988

Y1 - 1988

N2 - 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.

AB - 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.

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ER -

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

Abstract

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