title: Sequence specificity in the recognition of the epidermal growth factor receptor by the abl Src homology 2 domain.
abstract: The transforming activity of the abl gene product requires a functional src homology 2 (SH2) domain. An assay was developed to evaluate this function by examining binding of a bacterially-expressed abl SH2 domain to the activated EGF receptor, used as a surrogate tyrosine phosphorylated protein. The sequence specificity of this interaction has been explored with a series of point mutants of EGF receptor. Analysis of equilibrium binding reveals that substitution of Tyr1086 for Phe in the EGF receptor produced a 10-fold reduced affinity for abl SH2 domain binding as compared to the wildtype receptor. Moreover, a phosphorylated peptide modeled on the sequences surrounding Tyr1086 specifically inhibits abl SH2 binding, with an IC50 of approximately 10 microM. Evaluation of a series of additional peptides, modeled on the Tyr1086 sequence, revealed that the carboxy terminal residues directly next to the phosphotyrosine were particularly critical to this binding. Molecular modeling studies of the pTyr1086 peptide revealed the potential hydrophobic, ionic and hydrogen bonding interactions involved in the functions of the abl SH2 domain.
authors.affiliation: Department of Physiology, University of Michigan School of Medicine, Ann Arbor 48109.
authors.forename: G
authors.initials: G
authors.lastname: Zhu
authors.forename: S J
authors.initials: SJ
authors.lastname: Decker
authors.forename: D
authors.initials: D
authors.lastname: Maclean
authors.forename: D J
authors.initials: DJ
authors.lastname: McNamara
authors.forename: J
authors.initials: J
authors.lastname: Singh
authors.forename: T K
authors.initials: TK
authors.lastname: Sawyer
authors.forename: A R
authors.initials: AR
authors.lastname: Saltiel
mesh_terms: D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D066246:ErbB Receptors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017354:Point Mutation; D011487:Protein Conformation; D016315:Proto-Oncogene Proteins c-abl / Q000737:chemistry / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D066246:ErbB Receptors; D016315:Proto-Oncogene Proteins c-abl