Purification and properties of extracellular signal-regulated kinase 1, an insulin-stimulated microtubule-associated protein 2 kinase

Teri G. Boulton, Jill S. Gregory, Melanie H. Cobb

Research output: Contribution to journalArticle

114 Citations (Scopus)

Abstract

In rat 1 fibroblasts, insulin has little or no stimulatory effect on the activities of either MAP2 protein kinase or ribosomal protein S6 kinase. In contrast, in rat 1 cells that overexpress the normal human insulin receptor (rat 1 HIRc B; McClain et al. (1987) J. Biol. Chem. 262, 14663-14671), insulin activates both MAP2 and S6 kinase activities close to 5-fold. A MAP2 kinase has been purified from insulin-treated rat l HIRc B cells over 6300-fold by chromatography on Q-Sepharose, phenyl-Sepharose, S-Sepharose, phosphocellulose, QAE-Sepharose, UltrogelAcA54, DEAE-cellulose, and a second Q-Sepharose. Its specific activity is approximately 0.8-1 μmol·min-1·mg-1 with MAP2 and 3 μmol·min-1·mg-1 with myelin basic protein. The enzyme preparation contains one major band of Mr = 43 000 upon SDS-polyacrylamide gel electrophoresis, which is immunoblotted by antibodies to phosphotyrosine. A sequence from the 43-kDa band led to the isolation of a cDNA encoding the enzyme, which we have named ERK1 for extracellular signal-regulated kinase (Boulton et al. (1990) Science 249, 64-67).

Original languageEnglish (US)
Pages (from-to)278-286
Number of pages9
JournalBiochemistry®
Volume30
Issue number1
StatePublished - 1991

Fingerprint

Mitogen-Activated Protein Kinase 3
Sepharose
Purification
Rats
Insulin
Ribosomal Protein S6 Kinases
DEAE-Cellulose
Phosphotyrosine
Myelin Basic Protein
Extracellular Signal-Regulated MAP Kinases
Enzymes
Fibroblasts
Chromatography
Electrophoresis
Protein Kinases
Polyacrylamide Gel Electrophoresis
B-Lymphocytes
Phosphotransferases
Complementary DNA
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and properties of extracellular signal-regulated kinase 1, an insulin-stimulated microtubule-associated protein 2 kinase. / Boulton, Teri G.; Gregory, Jill S.; Cobb, Melanie H.

In: Biochemistry®, Vol. 30, No. 1, 1991, p. 278-286.

Research output: Contribution to journalArticle

@article{a224d1fac80b496ca1fc25537378c25a,
title = "Purification and properties of extracellular signal-regulated kinase 1, an insulin-stimulated microtubule-associated protein 2 kinase",
abstract = "In rat 1 fibroblasts, insulin has little or no stimulatory effect on the activities of either MAP2 protein kinase or ribosomal protein S6 kinase. In contrast, in rat 1 cells that overexpress the normal human insulin receptor (rat 1 HIRc B; McClain et al. (1987) J. Biol. Chem. 262, 14663-14671), insulin activates both MAP2 and S6 kinase activities close to 5-fold. A MAP2 kinase has been purified from insulin-treated rat l HIRc B cells over 6300-fold by chromatography on Q-Sepharose, phenyl-Sepharose, S-Sepharose, phosphocellulose, QAE-Sepharose, UltrogelAcA54, DEAE-cellulose, and a second Q-Sepharose. Its specific activity is approximately 0.8-1 μmol·min-1·mg-1 with MAP2 and 3 μmol·min-1·mg-1 with myelin basic protein. The enzyme preparation contains one major band of Mr = 43 000 upon SDS-polyacrylamide gel electrophoresis, which is immunoblotted by antibodies to phosphotyrosine. A sequence from the 43-kDa band led to the isolation of a cDNA encoding the enzyme, which we have named ERK1 for extracellular signal-regulated kinase (Boulton et al. (1990) Science 249, 64-67).",
author = "Boulton, {Teri G.} and Gregory, {Jill S.} and Cobb, {Melanie H.}",
year = "1991",
language = "English (US)",
volume = "30",
pages = "278--286",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - Purification and properties of extracellular signal-regulated kinase 1, an insulin-stimulated microtubule-associated protein 2 kinase

AU - Boulton, Teri G.

AU - Gregory, Jill S.

AU - Cobb, Melanie H.

PY - 1991

Y1 - 1991

N2 - In rat 1 fibroblasts, insulin has little or no stimulatory effect on the activities of either MAP2 protein kinase or ribosomal protein S6 kinase. In contrast, in rat 1 cells that overexpress the normal human insulin receptor (rat 1 HIRc B; McClain et al. (1987) J. Biol. Chem. 262, 14663-14671), insulin activates both MAP2 and S6 kinase activities close to 5-fold. A MAP2 kinase has been purified from insulin-treated rat l HIRc B cells over 6300-fold by chromatography on Q-Sepharose, phenyl-Sepharose, S-Sepharose, phosphocellulose, QAE-Sepharose, UltrogelAcA54, DEAE-cellulose, and a second Q-Sepharose. Its specific activity is approximately 0.8-1 μmol·min-1·mg-1 with MAP2 and 3 μmol·min-1·mg-1 with myelin basic protein. The enzyme preparation contains one major band of Mr = 43 000 upon SDS-polyacrylamide gel electrophoresis, which is immunoblotted by antibodies to phosphotyrosine. A sequence from the 43-kDa band led to the isolation of a cDNA encoding the enzyme, which we have named ERK1 for extracellular signal-regulated kinase (Boulton et al. (1990) Science 249, 64-67).

AB - In rat 1 fibroblasts, insulin has little or no stimulatory effect on the activities of either MAP2 protein kinase or ribosomal protein S6 kinase. In contrast, in rat 1 cells that overexpress the normal human insulin receptor (rat 1 HIRc B; McClain et al. (1987) J. Biol. Chem. 262, 14663-14671), insulin activates both MAP2 and S6 kinase activities close to 5-fold. A MAP2 kinase has been purified from insulin-treated rat l HIRc B cells over 6300-fold by chromatography on Q-Sepharose, phenyl-Sepharose, S-Sepharose, phosphocellulose, QAE-Sepharose, UltrogelAcA54, DEAE-cellulose, and a second Q-Sepharose. Its specific activity is approximately 0.8-1 μmol·min-1·mg-1 with MAP2 and 3 μmol·min-1·mg-1 with myelin basic protein. The enzyme preparation contains one major band of Mr = 43 000 upon SDS-polyacrylamide gel electrophoresis, which is immunoblotted by antibodies to phosphotyrosine. A sequence from the 43-kDa band led to the isolation of a cDNA encoding the enzyme, which we have named ERK1 for extracellular signal-regulated kinase (Boulton et al. (1990) Science 249, 64-67).

UR - http://www.scopus.com/inward/record.url?scp=0025976722&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025976722&partnerID=8YFLogxK

M3 - Article

VL - 30

SP - 278

EP - 286

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 1

ER -