Selection of v-abl tyrosine kinase substrate sequences from randomized peptide and cellular proteomic libraries using mRNA display

Chem Biol. 2002 Feb;9(2):253-64. doi: 10.1016/s1074-5521(02)00098-4.

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.

MeSH terms

  • Amino Acid Sequence
  • Bone Marrow Cells
  • Humans
  • Molecular Sequence Data
  • Oncogene Proteins v-abl / metabolism*
  • Peptide Library*
  • Phosphorylation
  • Protein-Tyrosine Kinases / metabolism*
  • RNA, Messenger / metabolism*
  • Recombinant Fusion Proteins
  • Substrate Specificity

Substances

  • Oncogene Proteins v-abl
  • Peptide Library
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Protein-Tyrosine Kinases