title: Augmentation of the affinity of HLA class I-binding peptides lacking primary anchor residues by manipulation of the secondary anchor residues.
abstract: A direct binding assay has been used to investigate the effect of the secondary anchor residues on peptide binding to class I proteins of the major histocompatibility complex. Based on predictions from a previous chemometric approach, synthetic peptide analogues containing unnatural amino acids were synthesized and tested for B*2705 binding. Hydrophobic unnatural amino acids such as alpha-naphthyl- and cyclohexyl-alanine were found to be excellent substituents in the P3 secondary anchor position giving peptides with very high B*2705-binding affinity. The binding to B*2705 of peptides optimized for their secondary anchor residues, but lacking one of the P2 or P9 primary anchor residues was also investigated. Most such peptides did not bind, but one peptide, lacking the P2 Arg residue generally considered essential for binding to all B27 subtypes, was found to bind quite strongly. These findings demonstrate that peptide binding to class I proteins is due to a combination of all the anchor residues, which may be occupied also by unnatural amino acids-a necessary step towards the development of peptidic or non-peptidic antagonists for immunomodulation.
authors.affiliation: Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Rovero
authors.forename: S
authors.initials: S
authors.lastname: Viganò
authors.forename: S
authors.initials: S
authors.lastname: Pegoraro
authors.forename: R
authors.initials: R
authors.lastname: Revoltella
authors.forename: D
authors.initials: D
authors.lastname: Riganelli
authors.forename: D
authors.initials: D
authors.lastname: Fruci
authors.forename: G
authors.initials: G
authors.lastname: Greco
authors.forename: R H
authors.initials: RH
authors.lastname: Butler
authors.forename: N
authors.initials: N
authors.lastname: Tanigaki
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015796:HLA-B27 Antigen / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008958:Models, Molecular; D015394:Molecular Structure; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding
chemical_list: D015796:HLA-B27 Antigen; D009842:Oligopeptides