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PMID: 7935430 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Association of MEK1 with p21ras.GMPPNP is dependent on B-Raf.

Molecular and cellular biology ·Vol. 14 ·No. 11 ·1994-11-00 ·Pages 7153-62

Moodie SA, Paris MJ, Kolch W, Wolfman A

Abstract

We have previously reported that immobilized p21ras forms a GMPPNP-dependent complex with a MEK activity. Furthermore, the association of the MEK activity was found to be independent of the presence of Raf-1. We have extended those observations to show that MEK1 is the MEK activity previously described to associate with immobilized p21ras.GMPPNP. The association between MEK1 and immobilized p21ras.GMPPNP increased its specific activity towards p42MAPK. We detected the specific association of B-Raf with immobilized p21ras.GMPPNP. In contrast to Raf-1-immunodepleted lysates, preclearance of the cytosolic B-Raf significantly reduced, by 96%, the amount of MEK1 activity associated with immobilized p21ras.GMPPNP. The decrease in MEK1 activity correlated with complete loss in the binding of both B-Raf and MEK1 proteins with immobilized p21ras.GMPPNP. These data suggest that the p21ras.GMPPNP-dependent activation of MEK1 in brain extracts is dependent on the presence of the B-Raf protein kinase.

MeSH Terms
Amino Acid Sequence Animals Brain/metabolism Guanylyl Imidodiphosphate/metabolism In Vitro Techniques MAP Kinase Kinase 1 Mitogen-Activated Protein Kinase Kinases Molecular Sequence Data Protein Serine-Threonine Kinases/genetics,metabolism Protein-Tyrosine Kinases/metabolism Proto-Oncogene Proteins/genetics,metabolism Proto-Oncogene Proteins c-raf Proto-Oncogene Proteins p21(ras)/metabolism Rats Recombinant Proteins/metabolism
Chemicals
Proto-Oncogene Proteins Recombinant Proteins Guanylyl Imidodiphosphate Protein-Tyrosine Kinases Protein Serine-Threonine Kinases Proto-Oncogene Proteins c-raf MAP Kinase Kinase 1 Mitogen-Activated Protein Kinase Kinases Proto-Oncogene Proteins p21(ras)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Moodie S A
Department of Cell Biology, Cleveland Clinic Foundation, Ohio 44195.
Paris M J
Kolch W
Wolfman A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.11.7153-7162.1994
Published
1994-11-00
Pages
7153-62
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359249
Subset
IM
Grants
NIGMS NIH HHS · GM 41220 · United States
NIGMS NIH HHS · GM 49652 · United States
External Links
PubMed source DOI: 10.1128/mcb.14.11.7153-7162.1994 PMC Full Text View journal details
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