title: A quantitative assay to measure the interaction between immunogenic peptides and purified class I major histocompatibility complex molecules.
abstract: A direct and sensitive biochemical assay to measure the interaction in solution between peptides and affinity-purified major histocompatibility complex (MHC) class I molecules has been generated. Specific binding reflecting the known class I restriction of cytotoxic T cell responses was obtained. Adding an excess of beta 2-microglobulin (beta 2m) significantly increased the rate of peptide association, but it did not affect the rate of dissociation. Binding was complicated by a rapid and apparently irreversible loss of functional MHC class I at 37 degrees C which might limit the life span of empty MHC class I thereby preventing the inadvertent exchange of peptides at the target cell surface. All class I molecules tested bound peptides of the canonical octa- to nona-meric length. However, one class I molecule, Kk, also bound peptides, which were much longer suggesting that the preference of class I molecules for short epitopes is not absolute and may be caused by factors other than the peptide-MHC class I binding event itself.
authors.affiliation: Institute for Medical Microbiology and Immunology, Medical Faculty, University of Copenhagen, Denmark.
authors.forename: A C
authors.initials: AC
authors.lastname: Olsen
authors.forename: L O
authors.initials: LO
authors.lastname: Pedersen
authors.forename: A S
authors.initials: AS
authors.lastname: Hansen
authors.forename: M H
authors.initials: MH
authors.lastname: Nissen
authors.forename: M
authors.initials: M
authors.lastname: Olsen
authors.forename: P R
authors.initials: PR
authors.lastname: Hansen
authors.forename: A
authors.initials: A
authors.lastname: Holm
authors.forename: S
authors.initials: S
authors.lastname: Buus
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry* / Q000378:metabolism; D001667:Binding, Competitive; D002460:Cell Line; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D051379:Mice; D008806:Mice, Inbred AKR; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding; D013696:Temperature; D013997:Time Factors; D001613:beta 2-Microglobulin / Q000378:metabolism
chemical_list: D000956:Antigens, Viral; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D001613:beta 2-Microglobulin