An examination of dynamics crosstalk between SH2 and SH3 domains by hydrogen/deuterium exchange and mass spectrometry

Protein Sci. 2006 Jan;15(1):65-73. doi: 10.1110/ps.051782206. Epub 2005 Dec 1.

Abstract

The ability of proteins to regulate their own enzymatic activity can be facilitated by changes in structure or protein dynamics in response to external regulators. Because many proteins contain SH2 and SH3 domains, transmission of information between the domains is a potential method of allosteric regulation. To determine if ligand binding to one modular domain may alter structural dynamics in an adjacent domain, allowing potential transmission of information through the protein, we used hydrogen exchange and mass spectrometry to measure changes in protein dynamics in the SH3 and SH2 domains of hematopoietic cell kinase (Hck). Ligand binding to either domain had little or no effect on hydrogen exchange in the adjacent domain, suggesting that changes in protein structure or dynamics are not a means of SH2/SH3 crosstalk. Furthermore, ligands of varying affinity covalently attached to SH3/SH2 altered dynamics only in the domain to which they bind. Such results demonstrate that ligand binding may not structurally alter adjacent SH3/SH2 domains and implies that other aspects of protein architecture contribute to the multiple levels of regulation in proteins containing SH3 and SH2 domains.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Allosteric Site
  • Deuterium Exchange Measurement*
  • Humans
  • Ligands
  • Mass Spectrometry
  • Peptide Fragments / chemistry*
  • Peptide Fragments / metabolism
  • Peptide Fragments / physiology
  • Protein Binding
  • Proto-Oncogene Proteins c-hck / chemistry
  • Proto-Oncogene Proteins c-hck / metabolism
  • Proto-Oncogene Proteins c-hck / physiology
  • Thermodynamics
  • src Homology Domains / physiology*

Substances

  • Ligands
  • Peptide Fragments
  • Proto-Oncogene Proteins c-hck