SOLUTION STRUCTURES OF ONCOGENE SH DOMAINS

Project: Research project

Project Details

Description

The objective of this project is to obtain high resolution solution
structures of proteins containing the SH (Src-Homology) domains 2 and 3
using multiple dimensional NMR. These domains are found in many proteins
that are involved in signal transducing pathways and have been implicated
in the inter- and intra-molecular regulation of kinase and other enzyme
activities and in the oncogenic potential of these proteins. As well, one
SH2 domain has been demonstrated to be sufficient for the high affinity
binding of tyrosyl-phosporylated proteins and and another SH3 domain may
bind actin filaments of the cellular cytoskeleton. This study is proposed
in order to generate the first three dimensional structures of these novel
domains and to identify their intramolecular interactions and
phosphotyrosyl-containing protein binding sites. The immediate goal is to
proceed from circular dichroism and two-dimensional NMR studies already
completed with unlabelled protein to a complete assignment and secondary
structure analysis of a N15 labelled 107 residue protein containing the 80-
amino acid SH2 domain from c-abl protein. A high resolution tertiary
structure determination using interproton distance constraints and torsion
angles from C-13 and N-15 labeled protein would be generated. Amino acid
residues involved in binding phosphotyrosyl containing substrate would be
identified by NMR. Since the transforming activity of several oncogenes
has been traced to mutations in their SH2 domains, the three dimensional
structure of the SH2 domain of the c-abl proto-oncogene should provide a
structural explanation for their oncogenic activity, and may suggest
rational routes to therapies for related conditions.
StatusFinished
Effective start/end date2/1/923/31/10

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)