Mapping of a small phosphopeptide at the carboxyterminus of the viral myb protein by monoclonal antibodies.
Several myb-specific monoclonal antibodies were produced and their antigen recognition sites characterized using a series of bacterially expressed truncated myb proteins. The monoclonal antibodies were used for analysing the in vivo phosphorylation site of the oncogene protein from avian myeloblastosis virus (AMV), p48v-myb. The p48v-myb protein labeled metabolically with [32P]orthophosphate was isolated from the AMV-transformed chicken myeloblast cell line BM-2 by immunoaffinity chromatography. Phosphoamino acid analysis indicated that it was phosphorylated mainly on serine and to a lesser extent (less than 5%) on threonine residues. Indirect immunoprecipitation of phosphopeptides from trypsin-digested [32P]-labeled purified p48v-myb protein by use of the myb-specific monoclonal antibodies allowed the mapping of a small phosphopeptide at the carboxyterminus of p48v-myb.
Bading
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H
Max-Planck-Institut für Molekulare Genetik, Berlin, Federal Republic of Germany.
Beutler
C
C
Moelling
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K
D000818:Animals; D000911:Antibodies, Monoclonal; D001354:Avian Leukosis Virus / Q000235:genetics*; D009189:Avian Myeloblastosis Virus / Q000235:genetics*; D002460:Cell Line; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics; D020626:Oncogene Proteins v-myb; D009857:Oncogenes; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000032:analysis*; D010766:Phosphorylation; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D012191:Retroviridae Proteins / Q000032:analysis* / Q000302:isolation & purification; D012694:Serine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D020626:Oncogene Proteins v-myb; D010446:Peptide Fragments; D010748:Phosphopeptides; D011993:Recombinant Fusion Proteins; D012191:Retroviridae Proteins; D012694:Serine