Structural and kinetic characterization of the simplified SH3 domain FP1

Protein Sci. 2003 Apr;12(4):776-83. doi: 10.1110/ps.0238603.

Abstract

The simplified SH3 domain sequence, FP1, obtained in phage display selection experiments has an amino acid composition that is 95% Ile, Lys, Glu, Ala, Gly. Here we use NMR to investigate the tertiary structure of FP1. We find that the overall topology of FP1 resembles that of the src SH3 domain, the hydrogen-deuterium exchange and chemical shift perturbation profiles are similar to those of naturally occurring SH3 domains, and the (15)N relaxation rates are in the range of naturally occurring small proteins. Guided by the structure, we further simplify the FP1 sequence and compare the effects on folding kinetics of point mutations in FP1 and the wild-type src SH3 domain. The results suggest that the folding transition state of FP1 is similar to but somewhat less polarized than that of the wild-type src SH3 domain.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Mutation
  • src Homology Domains / genetics
  • src Homology Domains / physiology*