Selection of v-abl tyrosine kinase substrate sequences from randomized peptide and cellular proteomic libraries using mRNA display.
Methodologies for rapidly identifying cellular protein interactions resulting in posttranslational modification of one of the partners are lacking. Here, we select for substrates of the v-abl tyrosine kinase from two protein display libraries in which the protein is covalently linked to its encoding mRNA. Successive selection cycles from a randomized peptide library identified a consensus sequence closely matching that previously reported for the v-abl tyrosine kinase. Selections from a proteomic library derived from cellular mRNA identified several novel targets of v-abl, including a new member of a class of SH2 domain-containing adaptor proteins. Upon modification, several of the substrates obtained in these selections were found to be effective inhibitors of v-abl kinase activity in vitro. These experiments establish a novel method for identifying the substrates of tyrosine kinases from synthetic and cellular protein libraries.
Cujec
Thomas P
TP
Phylos, 128 Spring Street, Lexington, MA 02421, USA. tcujec@phylos.com
Medeiros
Patricia F
PF
Hammond
Phil
P
Rise
Cecil
C
Kreider
Brent L
BL
D000595:Amino Acid Sequence; D001854:Bone Marrow Cells; D006801:Humans; D008969:Molecular Sequence Data; D016374:Oncogene Proteins v-abl / Q000378:metabolism*; D019151:Peptide Library; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D013379:Substrate Specificity
D016374:Oncogene Proteins v-abl; D019151:Peptide Library; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D011505:Protein-Tyrosine Kinases