title: Selection of v-abl tyrosine kinase substrate sequences from randomized peptide and cellular proteomic libraries using mRNA display.
abstract: 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.
authors.affiliation: Phylos, 128 Spring Street, Lexington, MA 02421, USA. tcujec@phylos.com
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Cujec
authors.forename: Patricia F
authors.initials: PF
authors.lastname: Medeiros
authors.forename: Phil
authors.initials: P
authors.lastname: Hammond
authors.forename: Cecil
authors.initials: C
authors.lastname: Rise
authors.forename: Brent L
authors.initials: BL
authors.lastname: Kreider
mesh_terms: 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
chemical_list: D016374:Oncogene Proteins v-abl; D019151:Peptide Library; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D011505:Protein-Tyrosine Kinases