title: Epitope mapping of Ly-49G and G-like receptors: CK-1 antibody defines a polymorphic site of functional interaction with class I ligand.
abstract: Ly-49 receptors regulate mouse natural killer cell functions. Members of the polymorphic Ly-49 multigene family recognize specific alleles of major histocompatibility complex class I (MHC I) or MHC I-like proteins. Previous studies have provided insight into the nature of Ly-49A and -C interaction with their high-affinity MHC I ligands, H-2Dd and Kb, respectively. Unlike Ly-49C, recognition of MHC I by Ly-49A is regulated in part by residues within the beta4-beta5 loop of its ectodomain. Ly-49A and -G are within the same Ly-49 subfamily, and both receptors recognize Dd. However, there have been no studies that define specific sites on Ly-49G that mediate class I MHC recognition. The Ly-49G receptors of different inbred mouse strains can differ as a result of amino acid polymorphisms within their ectodomains. In this report, we have generated a novel antibody, CK-1, which recognizes Ly-49G(B6) and a Ly-49G(B6)-like receptor, Ly-49M(nonobese diabetic), but not Ly-49G(BALB/c). By exploiting the differences within ectodomains of C57BL/6 and BALB/c Ly-49G allele products, we identified epitopes recognized by the Ly-49G-specific antibodies CK-1 and Cwy-3, whose epitopes mapped within the beta4-beta5 loop and the beta1 strand, respectively, and were nonoverlapping. Although both antibodies specifically recognized the Ly-49G(B6) ectodomain, Cwy-3 was unable to block its interaction with MHC I, and CK-1 significantly inhibited it. The importance of residues within the beta4-beta5 loop in Ly-49G recognition demonstrates that its interaction with MHC I is similar to that of Ly-49A but not Ly-49C.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada, T6G 2S2.
authors.forename: Mohammed S
authors.initials: MS
authors.lastname: Osman
authors.forename: Elizabeth T
authors.initials: ET
authors.lastname: Silver
authors.forename: Jay C
authors.initials: JC
authors.lastname: Varghese
authors.forename: Chew Shun
authors.initials: CS
authors.lastname: Chang
authors.forename: Dong-Er
authors.initials: DE
authors.lastname: Gong
authors.forename: Gerald F
authors.initials: GF
authors.lastname: Audette
authors.forename: Bart
authors.initials: B
authors.lastname: Hazes
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Kane
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000950:Antigens, Ly / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D018604:Epitope Mapping; D005434:Flow Cytometry; D005805:Genes, MHC Class I / Q000276:immunology; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology*; D037181:Lectins, C-Type; D016131:Lymphocyte Subsets / Q000166:cytology / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D017434:Protein Structure, Tertiary; D055630:Receptors, NK Cell Lectin-Like; D016415:Sequence Alignment; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000950:Antigens, Ly; D037181:Lectins, C-Type; D055630:Receptors, NK Cell Lectin-Like