An insulin-stimulated ribosomal protein S6 kinase from rabbit liver

J. S. Gregory, T. G. Boulton, B. C. Sang, M. H. Cobb

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

In this report we describe an activated form of S6 protein kinase in rabbits treated acutely with insulin. The major insulin-stimulated activity in rabbit liver is increased 2- to 5-fold compared to material from untreated animals based on DEAE-cellulose profiles. The activity observed in DEAE-cellulose fractions can be separated into a major and minor peak, each having very similar chromatographic behavior. Chromatography on DEAE-cellulose, S-Sepharose, heptyl-Sepharose, heparin-agarose, and Mono Q results in greater than 20,000-fold purification of the insulin-stimulated enzyme with a 12% recovery. The stimulated activity has chromatographic properties similar to an S6 protein kinase studied previously in 3T3-L1 cells (Cobb, M.H. (1986) J. Biol. Chem. 261, 12994-12999) and other systems. The enzyme purified from insulin-treated animals contains a major band that migrates in sodium dodecyl sulfate-polyacrylamide gels with M(r) ~ 70,000; this band also appears in the control preparation. Treatment of the insulin-stimulated S6 kinase with the catalytic subunit of phosphatase 2a reduces its activity by 97%. The activity of the inactivated S6 kinase is stimulated nearly 5-fold by a 15-min preincubation with partially purified insulin-stimulated microtubule-associated protein-2 kinase.

Original languageEnglish (US)
Pages (from-to)18397-18401
Number of pages5
JournalJournal of Biological Chemistry
Volume264
Issue number31
StatePublished - 1989

Fingerprint

Ribosomal Protein S6 Kinases
Liver
Insulin
Rabbits
DEAE-Cellulose
Sepharose
Protein Kinases
Animals
3T3-L1 Cells
DEAE-Cellulose Chromatography
Mitogen-Activated Protein Kinase 3
Enzymes
Chromatography
Phosphoric Monoester Hydrolases
Sodium Dodecyl Sulfate
Purification
Catalytic Domain
Recovery

ASJC Scopus subject areas

  • Biochemistry

Cite this

Gregory, J. S., Boulton, T. G., Sang, B. C., & Cobb, M. H. (1989). An insulin-stimulated ribosomal protein S6 kinase from rabbit liver. Journal of Biological Chemistry, 264(31), 18397-18401.

An insulin-stimulated ribosomal protein S6 kinase from rabbit liver. / Gregory, J. S.; Boulton, T. G.; Sang, B. C.; Cobb, M. H.

In: Journal of Biological Chemistry, Vol. 264, No. 31, 1989, p. 18397-18401.

Research output: Contribution to journalArticle

Gregory, JS, Boulton, TG, Sang, BC & Cobb, MH 1989, 'An insulin-stimulated ribosomal protein S6 kinase from rabbit liver', Journal of Biological Chemistry, vol. 264, no. 31, pp. 18397-18401.
Gregory, J. S. ; Boulton, T. G. ; Sang, B. C. ; Cobb, M. H. / An insulin-stimulated ribosomal protein S6 kinase from rabbit liver. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 31. pp. 18397-18401.
@article{2382389cedb2462ab134a55b52ee6612,
title = "An insulin-stimulated ribosomal protein S6 kinase from rabbit liver",
abstract = "In this report we describe an activated form of S6 protein kinase in rabbits treated acutely with insulin. The major insulin-stimulated activity in rabbit liver is increased 2- to 5-fold compared to material from untreated animals based on DEAE-cellulose profiles. The activity observed in DEAE-cellulose fractions can be separated into a major and minor peak, each having very similar chromatographic behavior. Chromatography on DEAE-cellulose, S-Sepharose, heptyl-Sepharose, heparin-agarose, and Mono Q results in greater than 20,000-fold purification of the insulin-stimulated enzyme with a 12{\%} recovery. The stimulated activity has chromatographic properties similar to an S6 protein kinase studied previously in 3T3-L1 cells (Cobb, M.H. (1986) J. Biol. Chem. 261, 12994-12999) and other systems. The enzyme purified from insulin-treated animals contains a major band that migrates in sodium dodecyl sulfate-polyacrylamide gels with M(r) ~ 70,000; this band also appears in the control preparation. Treatment of the insulin-stimulated S6 kinase with the catalytic subunit of phosphatase 2a reduces its activity by 97{\%}. The activity of the inactivated S6 kinase is stimulated nearly 5-fold by a 15-min preincubation with partially purified insulin-stimulated microtubule-associated protein-2 kinase.",
author = "Gregory, {J. S.} and Boulton, {T. G.} and Sang, {B. C.} and Cobb, {M. H.}",
year = "1989",
language = "English (US)",
volume = "264",
pages = "18397--18401",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "31",

}

TY - JOUR

T1 - An insulin-stimulated ribosomal protein S6 kinase from rabbit liver

AU - Gregory, J. S.

AU - Boulton, T. G.

AU - Sang, B. C.

AU - Cobb, M. H.

PY - 1989

Y1 - 1989

N2 - In this report we describe an activated form of S6 protein kinase in rabbits treated acutely with insulin. The major insulin-stimulated activity in rabbit liver is increased 2- to 5-fold compared to material from untreated animals based on DEAE-cellulose profiles. The activity observed in DEAE-cellulose fractions can be separated into a major and minor peak, each having very similar chromatographic behavior. Chromatography on DEAE-cellulose, S-Sepharose, heptyl-Sepharose, heparin-agarose, and Mono Q results in greater than 20,000-fold purification of the insulin-stimulated enzyme with a 12% recovery. The stimulated activity has chromatographic properties similar to an S6 protein kinase studied previously in 3T3-L1 cells (Cobb, M.H. (1986) J. Biol. Chem. 261, 12994-12999) and other systems. The enzyme purified from insulin-treated animals contains a major band that migrates in sodium dodecyl sulfate-polyacrylamide gels with M(r) ~ 70,000; this band also appears in the control preparation. Treatment of the insulin-stimulated S6 kinase with the catalytic subunit of phosphatase 2a reduces its activity by 97%. The activity of the inactivated S6 kinase is stimulated nearly 5-fold by a 15-min preincubation with partially purified insulin-stimulated microtubule-associated protein-2 kinase.

AB - In this report we describe an activated form of S6 protein kinase in rabbits treated acutely with insulin. The major insulin-stimulated activity in rabbit liver is increased 2- to 5-fold compared to material from untreated animals based on DEAE-cellulose profiles. The activity observed in DEAE-cellulose fractions can be separated into a major and minor peak, each having very similar chromatographic behavior. Chromatography on DEAE-cellulose, S-Sepharose, heptyl-Sepharose, heparin-agarose, and Mono Q results in greater than 20,000-fold purification of the insulin-stimulated enzyme with a 12% recovery. The stimulated activity has chromatographic properties similar to an S6 protein kinase studied previously in 3T3-L1 cells (Cobb, M.H. (1986) J. Biol. Chem. 261, 12994-12999) and other systems. The enzyme purified from insulin-treated animals contains a major band that migrates in sodium dodecyl sulfate-polyacrylamide gels with M(r) ~ 70,000; this band also appears in the control preparation. Treatment of the insulin-stimulated S6 kinase with the catalytic subunit of phosphatase 2a reduces its activity by 97%. The activity of the inactivated S6 kinase is stimulated nearly 5-fold by a 15-min preincubation with partially purified insulin-stimulated microtubule-associated protein-2 kinase.

UR - http://www.scopus.com/inward/record.url?scp=0024464069&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024464069&partnerID=8YFLogxK

M3 - Article

VL - 264

SP - 18397

EP - 18401

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 31

ER -