Abstract
The Raf kinase inhibitory protein (RKIP) binds to Raf-1 interfering with binding of the MEK substrate and potentially also Raf-1 activation. In response to mitogen stimulation RKIP dissociates from Raf-1 and later re-associates. Here, using a combination of mutational approaches, biochemical studies, peptide arrays and plasmon surface resonance (BIAcore), we fine map and characterize a minimal 24 amino acid long RKIP binding domain in the Raf-1 N-region, which consists of constitutive elements at both flanks and a center element that is regulated by phosphorylation and enhances the re-binding of RKIP to Raf-1 in the later phase of mitogen stimulation.
Publication types
-
Research Support, N.I.H., Extramural
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
COS Cells
-
Chlorocebus aethiops
-
Enzyme Activation / drug effects
-
Enzyme Activation / genetics
-
Humans
-
MAP Kinase Kinase Kinases / genetics
-
MAP Kinase Kinase Kinases / metabolism
-
Mitogens / pharmacology
-
Phosphatidylethanolamine Binding Protein / genetics
-
Phosphatidylethanolamine Binding Protein / metabolism*
-
Phosphorylation
-
Protein Processing, Post-Translational / drug effects
-
Protein Processing, Post-Translational / genetics
-
Protein Structure, Tertiary / genetics
-
Proto-Oncogene Proteins c-raf / genetics
-
Proto-Oncogene Proteins c-raf / metabolism*
-
Surface Plasmon Resonance / methods
Substances
-
Mitogens
-
PEBP1 protein, human
-
Phosphatidylethanolamine Binding Protein
-
Proto-Oncogene Proteins c-raf
-
MAP Kinase Kinase Kinases