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PMID: 2402489 Published · ppublish English Journal Article

Identical Mr 70,000 S6 kinase is activated biphasically by epidermal growth factor: a phosphopeptide that characterizes the late phase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 18 ·1990-09-00 ·Pages 7040-4

Susa M, Thomas G

Abstract

Mitogenic stimulation of quiescent mouse 3T3 cells with epidermal growth factor leads to biphasic S6 kinase activation. The kinases present in both phases of the response have been purified from 32P-labeled cells and shown to contain a phosphoprotein of equivalent Mr 70,000. Chromatographic analysis of the purified S6 kinases on a Mono Q column reveals that (i) all 32P-labeled protein coelutes with S6 kinase activity, (ii) only those fractions containing S6 kinase autophosphorylate, (iii) autophosphorylation is restricted to a single Mr 70,000 protein, and (iv) the extent of autophosphorylation directly parallels the degree of S6 kinase activation. Analysis of the two autophosphorylated S6 kinases by two-dimensional tryptic phosphopeptide mapping indicates that they are the same protein. Both in vivo 32P-labeled S6 kinases contain phosphoserine and phosphothreonine but no detectable phosphotyrosine. Two-dimensional tryptic peptide maps of the in vivo 32P-labeled S6 kinases are essentially identical, except for a single qualitative change in the late-phase S6 kinase.

MeSH Terms
Animals Cell Line Enzyme Activation Epidermal Growth Factor/pharmacology Kinetics Mice Molecular Weight Peptide Mapping Phosphates/metabolism Phosphopeptides/analysis Phosphorylation Protein Kinases/metabolism Ribosomal Protein S6 Kinases Trypsin
Chemicals
Phosphates Phosphopeptides Epidermal Growth Factor Protein Kinases Ribosomal Protein S6 Kinases Trypsin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Susa M
Friedrich Miescher Institute, Basel, Switzerland.
Thomas G
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43 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.18.7040
Published
1990-09-00
Pages
7040-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54678
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.87.18.7040 PMC Full Text View journal details
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