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PMID: 2426575 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Restriction of the in vitro and in vivo tyrosine protein kinase activities of pp60c-src relative to pp60v-src.

Molecular and cellular biology ·Vol. 5 ·No. 10 ·1985-10-00 ·Pages 2753-63

Coussens PM, Cooper JA, Hunter T, Shalloway D

Abstract

The tyrosine protein kinase activities of pp60c-src and pp60v-src were compared. The activities were qualitatively similar in vitro when the src proteins were bound in an immune complex with monoclonal antibody; both proteins utilized either ATP or GTP as phosphate donors, preferred Mn2+ to Mg2+, and had similar exogenous substrate specificities. The specific activity of pp60c-src was about 10-fold lower than that of pp60v-src for exogenous substrate phosphorylation but was only 1.1- to 2-fold lower than that of pp60v-src for autophosphorylation. Six glycolytic enzymes, including three not previously identified as substrates for pp60src phosphorylation, were phosphorylated by both pp60c-src and pp60v-src. Levels of pp60c-src fourfold higher than the amount of pp60v-src in src-plasmid-transformed cells did not detectably alter the level of phosphotyrosine in cellular proteins, but increasing the expression of pp60c-src another twofold (which induces cells to form foci in monolayer culture (P.J. Johnson, P.M. Coussens, A.V. Danko, and D. Shalloway, Mol. Cell. Biol. 5:1073-1083, 1985) resulted in a threefold increase in the level of cellular protein phosphotyrosine. Immunoprecipitation and analysis of the alkali-stable phosphoproteins by two-dimensional electrophoresis showed that, in contrast to pp60v-src-transformed cells, pp36 and enolase are only weakly phosphorylated in these high-level pp60c-src overexpresser cells. Even allowing for the in vitro differences in specific activities of phosphorylation, these results suggest that the pp60c-src tyrosine protein phosphorylating activity may be restricted relative to that of pp60v-src by additional in vivo mechanisms.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Cell Line Chickens Guanosine Triphosphate/metabolism Immunoglobulin G/metabolism In Vitro Techniques Isoelectric Point Mice Molecular Weight Oncogene Protein pp60(v-src) Phosphorylation Phosphotyrosine Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins/metabolism Proto-Oncogene Proteins pp60(c-src) Retroviridae Proteins/metabolism Substrate Specificity Tyrosine/analogs & derivatives,metabolism
Chemicals
Immunoglobulin G Proto-Oncogene Proteins Retroviridae Proteins Phosphotyrosine Tyrosine Guanosine Triphosphate Adenosine Triphosphate Protein-Tyrosine Kinases Oncogene Protein pp60(v-src) Proto-Oncogene Proteins pp60(c-src)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Coussens P M
Cooper J A
Hunter T
Shalloway D
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50 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.5.10.2753-2763.1985
Published
1985-10-00
Pages
2753-63
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367013
Subset
IM
Grants
NCI NIH HHS · CA17096 · United States
NCI NIH HHS · CA28485 · United States
NCI NIH HHS · CA32317 · United States
External Links
PubMed source DOI: 10.1128/mcb.5.10.2753-2763.1985 PMC Full Text View journal details
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