Crystallization and preliminary X-ray crystallographic characterization of a public CMV-specific TCR in complex with its cognate antigen.
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.
Reiser
Jean Baptiste
JB
Institut de Biologie Structurale Jean-Pierre Ebel, UMR, Grenoble, France.
Legoux
François
F
Machillot
Paul
P
Debeaupuis
Emilie
E
Le Moullac-Vaydie
Béatrice
B
Chouquet
Anne
A
Saulquin
Xavier
X
Bonneville
Marc
M
Housset
Dominique
D
D000956:Antigens, Viral / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D003460:Crystallization; D018360:Crystallography, X-Ray; D003587:Cytomegalovirus / Q000737:chemistry* / Q000276:immunology; D015789:HLA-A2 Antigen / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry* / Q000276:immunology; D014961:X-Ray Diffraction
D000956:Antigens, Viral; D015789:HLA-A2 Antigen; D016056:Immunodominant Epitopes; D010455:Peptides; D011948:Receptors, Antigen, T-Cell