Probing the ligand-binding specificity and analyzing the folding state of SPOT-synthesized FBP28 WW domain variants

Chembiochem. 2006 May;7(5):780-8. doi: 10.1002/cbic.200500408.

Abstract

The WW domains are known as the smallest naturally occurring, monomeric, triple-stranded, antiparallel beta-sheet domains. Hence, we chose the FBP28 WW domain as a model to investigate the stability of the beta-sheet structure at the amino acid level in the context of its function (ligand binding). The structure-function relationship was investigated through a complete substitution analysis of the FBP28 WW domain, with variants synthesized as a cellulose-bound peptide array. The functionality of the FBP28 WW domain variants was examined by probing the peptide array for ligand binding. In addition, selected FBP28 WW domain variants were investigated by CD measurements to determine the stability of the antiparallel beta-sheet structure. We discuss the correlation between structure stability and functionality for the FBP28 WW domain, as well as the effect of ligand-induced structure stabilization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / chemical synthesis
  • Carrier Proteins / chemistry*
  • Cellulose / chemistry
  • Circular Dichroism
  • Humans
  • Ligands
  • Peptides / chemical synthesis
  • Peptides / chemistry*
  • Protein Array Analysis / methods
  • Protein Binding
  • Protein Folding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Sensitivity and Specificity
  • Structure-Activity Relationship

Substances

  • Carrier Proteins
  • Ligands
  • Peptides
  • formin binding protein 28, human
  • Cellulose