Crystallization and preliminary X-ray crystallographic characterization of a public CMV-specific TCR in complex with its cognate antigen

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2009 Nov 1;65(Pt 11):1157-61. doi: 10.1107/S1744309109037890. Epub 2009 Oct 30.

Abstract

The T-cell response to human cytomegalovirus is characterized by a dramatic reduction of clonal diversity in patients undergoing chronic inflammation or immunodepression. In order to check whether all the selected high-avidity T-cell clones recognize the immunodominant pp65 peptide antigen pp65(495-503) (NLVPMVATV) presented by the major histocompatibility complex (MHC) molecule HLA-A2 in a similar manner, several public high-affinity T-cell receptors (TCRs) specific for the pp65(495-503)-HLA-A2 complex have been investigated. Expression, purification and crystallization were performed and preliminary crystallographic data were collected to 4.7 angstrom resolution for the RA15 TCR in complex with the pp65(495-503)-HLA-A2 complex. Comparison of the RA15-pp65(495-503)-HLA-A2 complex molecular-replacement solution with the structure of another high-affinity pp65(495-503)-HLA-A2-specific TCR, RA14, shows a shared docking mode, indicating that the clonal focusing could be accompanied by the selection of a most favoured peptide-readout mode. However, the position of the RA15 V beta domain is significantly shifted, suggesting a different interatomic interaction network.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Viral / chemistry*
  • Antigens, Viral / genetics
  • Antigens, Viral / immunology
  • Crystallization
  • Crystallography, X-Ray
  • Cytomegalovirus / chemistry*
  • Cytomegalovirus / immunology
  • HLA-A2 Antigen / chemistry
  • HLA-A2 Antigen / genetics
  • HLA-A2 Antigen / immunology
  • Humans
  • Immunodominant Epitopes / chemistry*
  • Immunodominant Epitopes / genetics
  • Immunodominant Epitopes / immunology
  • Models, Molecular
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / genetics
  • Peptides / immunology
  • Protein Conformation
  • Receptors, Antigen, T-Cell / chemistry*
  • Receptors, Antigen, T-Cell / immunology
  • X-Ray Diffraction

Substances

  • Antigens, Viral
  • HLA-A2 Antigen
  • Immunodominant Epitopes
  • Peptides
  • Receptors, Antigen, T-Cell