Probing structure and function of the raf protein kinase domain with monoclonal antibodies.
Five monoclonal antibodies were generated against the raf kinase domain. All antibodies react with different isozymes of the raf family, as well as Raf proteins from different species, albeit with differential affinities. Epitope mapping showed all five epitopes clustered in the vicinity of the conserved APE sequence. Although thought to be an essential part of the catalytic site, antibody binding to that domain does not affect kinase activity in vitro or the capability to specifically associate with other cellular proteins. Based on a detailed dissection of the epitopes, a comparative analysis of secondary structure predictions indicates a common structural motif in that region, which is highly conserved amongst protein kinases of the serine/threonine as well as the tyrosine class.
Kolch
W
W
Laboratory of Viral Carcinogenesis, National Cancer Institute, Frederick Cancer Research Facility, Maryland 21701.
Weissinger
E
E
Mischak
H
H
Troppmair
J
J
Showalter
S D
SD
Lloyd
P
P
Heidecker
G
G
Rapp
U R
UR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D002874:Chromosome Mapping; D000939:Epitopes / Q000276:immunology; D006825:Hybridomas / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009101:Multiple Myeloma / Q000276:immunology / Q000473:pathology; D011233:Precipitin Tests; D011487:Protein Conformation; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D019908:Proto-Oncogene Proteins c-raf
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011518:Proto-Oncogene Proteins; D011494:Protein Kinases; D019908:Proto-Oncogene Proteins c-raf