title: Rigidification of the alpha2 helix of an MHC class I molecule by a valine to proline mutation in position 165 does not prevent peptide-specific antigen presentation.
abstract: Although classical MHC class I glycoproteins bind peptide Ags for display at the cell surface, some MHC class I-related molecules such as the neonatal Fc receptor (FcRn) execute their function without binding peptide ligands. The three-dimensional structure of the FcRn suggested that a substitution of the conserved valine at position 165 of the alpha2 helix by proline contributed to a kink in the position of this helix relative to the alpha1 helix, and resulted in closing of the potential peptide-binding cleft. To test the contribution of proline 165 to the occlusion of the cleft and the binding of potential antigenic peptides, we introduced this mutation into the classical murine MHC class I molecule, H-2Dd, and characterized the ability of such a mutant to present peptide Ags to either a peptide-specific, H-2Dd-restricted T cell hybridoma (B4.2.3), or an allospecific, peptide-dependent, T cell hybridoma (3DT52.5.8). We show that the V165P mutation, expressed at the cell surface either in H-2Dd or in a single chain membrane version of H-2Dd, fails to eliminate recognition of the peptide/MHC complexes by two different T cells. Evaluation of a panel of synthetic substituted peptides suggests that subtle differences in the fine specificity of presentation can be discerned. Thus, the proline substitution at position 165 of FcRn and some other class I-like molecules is not the sole cause of the lack of peptide presentation.
authors.affiliation: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1892, USA.
authors.forename: D
authors.initials: D
authors.lastname: Plaksin
authors.forename: K
authors.initials: K
authors.lastname: Polakova
authors.forename: M G
authors.initials: MG
authors.lastname: Mage
authors.forename: D H
authors.initials: DH
authors.lastname: Margulies
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D006825:Hybridomas; D007739:L Cells; D051379:Mice; D010455:Peptides / Q000276:immunology; D017354:Point Mutation; D011392:Proline / Q000235:genetics; D017510:Protein Folding; D013601:T-Lymphocytes; D014633:Valine / Q000235:genetics
chemical_list: D015395:Histocompatibility Antigens Class I; D010455:Peptides; D011392:Proline; D014633:Valine