The Aminoterminus of c-Raf-1 Binds a Protein Kinase Phosphorylating Ser259

• Published in: Biochemical and biophysical research communications Vol. 204; no. 2; pp. 841 - 848
• Main Authors: Beimling, P., Niehof, M., Radziwill, G., Moelling, K.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 28.10.1994
• Subjects: Amino Acid Sequence; Glutathione Transferase - metabolism; Humans; Molecular Sequence Data; Phosphorylation; Protein Binding; Protein Kinases - metabolism; Protein-Serine-Threonine Kinases - chemistry; Protein-Serine-Threonine Kinases - metabolism; Proto-Oncogene Proteins - chemistry; Proto-Oncogene Proteins - metabolism; Proto-Oncogene Proteins c-raf; Recombinant Fusion Proteins - metabolism; Serine - metabolism; Tumor Cells, Cultured
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1994.2536

The kinase negative aminoterminal domain of c-Raf-1 expressed as glutathione S-transferase fusion protein was phosphorylated in vitro after treatment with lysates from A431 cells and subsequent in vitro protein kinase assay. This phosphorylation was independent of stimulation of the cells with EGF; it occurred exclusively on serine and was mapped to Ser259. The identical site of c-Raf-1 was phosphorylated in A431 cells by metabolic labelling in vivo. The kinase binding domain was mapped by various GST-Raf deletion mutants to c-Raf-1 aminoacid residues 181 to 255.