Abstract
Recombinant Mek1 and Raf-1 proteins produced in Sf9 cells undergo a tight association both in vivo and in vitro, which apparently does not depend on additional factors or the kinase activity of Mek1 or Raf-1. The complex can be disrupted by two polyclonal antibodies raised against Raf-1 peptides. Coinfection with Raf-1 activates Mek1 > 150-fold, and coinfection with Raf-1 and Mek1 activates Erk1 approximately 90-fold. The activation of Mek1 by Raf-1 involves only serine phosphorylation, which is directly proportional to the extent of Mek1 activation. Phosphopeptide maps suggest a single Raf-1 phosphorylation site on mek1.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Enzyme Activation
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In Vitro Techniques
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MAP Kinase Kinase 1
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Macromolecular Substances
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Mice
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Mitogen-Activated Protein Kinase Kinases*
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Phosphorylation
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Phosphoserine / metabolism
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Protein Binding
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Protein Serine-Threonine Kinases / metabolism*
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Protein-Tyrosine Kinases / metabolism*
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Proto-Oncogene Proteins / metabolism*
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Proto-Oncogene Proteins c-raf
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Recombinant Proteins
Substances
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Macromolecular Substances
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Proto-Oncogene Proteins
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Recombinant Proteins
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Phosphoserine
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Protein-Tyrosine Kinases
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Protein Serine-Threonine Kinases
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Proto-Oncogene Proteins c-raf
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MAP Kinase Kinase 1
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Map2k1 protein, mouse
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Mitogen-Activated Protein Kinase Kinases