title: Expression, refolding, crystallization and preliminary crystallographic study of MHC H-2Kk complexed with octapeptides and nonapeptides.
abstract: Major histocompatibility complex (MHC) molecules are heterodimeric cell-surface receptors that play a crucial role in the cellular immune response by presenting epitope peptides to T-cell antigen receptors (TCR). Although the structural basis of the peptide-MHC binding mechanism is becoming better understood, it is still difficult to predict a binding mode for an MHC of unknown structure. Therefore, as the first stage of a TCR-MHC interaction study, the crystal structures of the mouse H-2K(k) molecule in complex with both an octapeptide from Influenza A virus and a nonapeptide from simian virus SV40 were solved. Here, the expression, refolding, purification and crystallization of the two complexes are reported. For the H-2K(k)-HA(259-266) complex, crystals were obtained via an extensive screen using a nanodrop-dispensing robot and diffracted to 2.5 A resolution. For the H-2K(k)-SV40(560-568) complex, microscopic needles were initially obtained and their size was improved by macroseeding and a stepwise increase in precipitant concentration. Diffraction data to a resolution of 3.0 A were collected at a synchrotron facility.
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, UMR 6102, Parc Scientifique et Technologique de Luminy - Case 906, 13288 Marseille, France. kellenberger@ciml.univ-mrs.fr
authors.forename: Christine
authors.initials: C
authors.lastname: Kellenberger
authors.forename: Sophie
authors.initials: S
authors.lastname: Porciero
authors.forename: Alain
authors.initials: A
authors.lastname: Roussel
mesh_terms: D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006183:H-2 Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009980:Influenza A virus / Q000737:chemistry; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D020673:Protein Renaturation; D013539:Simian virus 40 / Q000737:chemistry
chemical_list: D006183:H-2 Antigens; D010446:Peptide Fragments