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

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

Research output: Contribution to journal › Article › peer-review

428 Scopus citations

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
State	Published - Jan 1988
Externally published	Yes

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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 J. Resink and Peter Stralfors and \{Lim Tung\}, \{H. Y.\}",

year = "1988",

month = jan,

doi = "10.1016/0076-6879(88)59039-0",

language = "English",

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journal = "Methods in Enzymology",

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TY - JOUR

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

AU - Cohen, Philip

AU - Alemany, Susana

AU - Therese, Brian A.

AU - Resink, J.

AU - Stralfors, Peter

AU - Lim Tung, H. Y.

PY - 1988/1

Y1 - 1988/1

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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U2 - 10.1016/0076-6879(88)59039-0

DO - 10.1016/0076-6879(88)59039-0

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AN - SCOPUS:0023737187

SN - 0076-6879

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SP - 390

EP - 408

JO - Methods in Enzymology

JF - Methods in Enzymology

IS - C

ER -

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

Abstract

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