title: HLA class I binding motifs derived from random peptide libraries differ at the COOH terminus from those of eluted peptides.
abstract: Recombinant HLA-A2, HLA-B8, or HLA-B53 heavy chain produced in Escherichia coli was combined with recombinant beta2-microglobulin (beta2m) and a pool of randomly synthesised nonamer peptides. This mixture was allowed to refold to form stable major histocompatability complex (MHC) class I complexes, which were then purified by gel filtration chromatography. The peptides bound to the MHC class I molecules were subsequently eluted and sequenced as a pool. Peptide binding motifs for these three MHC class I molecules were derived and compared with previously described motifs derived from analysis of naturally processed peptides eluted from the surface of cells. This comparison indicated that the peptides bound by the recombinant MHC class I molecules showed a similar motif to naturally processed and presented peptides, with the exception of the peptide COOH terminus. Whereas the motifs derived from naturally processed peptides eluted from HLA-A2 and HLA-B8 indicated a strong preference for hydrophobic amino acids at the COOH terminus, this preference was not observed in our studies. We propose that this difference reflects the effects of processing or transport on the peptide repertoire available for binding to MHC class I molecules in vivo.
authors.affiliation: Institute of Molecular Medicine, Nuffield Department of Medicine, University of Oxford, Headington, United Kingdom.
authors.forename: M P
authors.initials: MP
authors.lastname: Davenport
authors.forename: K J
authors.initials: KJ
authors.lastname: Smith
authors.forename: D
authors.initials: D
authors.lastname: Barouch
authors.forename: S W
authors.initials: SW
authors.lastname: Reid
authors.forename: W M
authors.initials: WM
authors.lastname: Bodnar
authors.forename: A C
authors.initials: AC
authors.lastname: Willis
authors.forename: D F
authors.initials: DF
authors.lastname: Hunt
authors.forename: A V
authors.initials: AV
authors.lastname: Hill
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006680:HLA Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library
chemical_list: D006680:HLA Antigens; D019151:Peptide Library