title: Somatic mutations to arginine residues affect the binding of human monoclonal antibodies to DNA, histones, SmD and Ro antigen.
abstract: Autoantibodies to a wide variety of antigens are associated with systemic lupus erythematosus (SLE). Antibodies to double-stranded DNA (anti-dsDNA) are thought to be particularly closely related to tissue damage and disease activity in SLE. Autoantibodies to histones, Sm and Ro are found in patients with SLE, but their role in pathogenesis is unclear. Using a transient expression system, we previously showed that particular sequence motifs in CDRs of light chains derived from the human Vlambda gene 2a2 are very important in determining their ability to form a DNA-binding site, when paired with the heavy chain of the human monoclonal anti-dsDNA antibody B3. These motifs are often sites of somatic mutation and/or contain arginine residues. In the experiments reported in this paper, the same expression system was used to show that these CDR motifs also affect binding to histones, Ro antigen and Sm antigen, but that binding to different antigens is affected in diverse ways by particular changes in the sequence of the CDRs. The heavy chain also plays a role in binding to these antigens. Pairing of the same range of 11 2a2 derived light chains with the heavy chain of a different anti-DNA antibody, 33.H11, gave reduced ability to bind DNA in comparison with the results obtained using the B3 heavy chain. Computer-generated models of the three-dimensional structures of these heavy/light chain combinations were used to define the positions occupied by the important sequence motifs at the binding sites of these antibodies, and to explain the different effects exerted by arginine residues at different positions in the light chains.
authors.affiliation: Centre for Rheumatology/Bloomsbury Rheumatology Unit, Division of Medicine, University College, London, UK.
authors.forename: Joanna
authors.initials: J
authors.lastname: Haley
authors.forename: Lesley J
authors.initials: LJ
authors.lastname: Mason
authors.forename: Sylvia
authors.initials: S
authors.lastname: Nagl
authors.forename: Ian
authors.initials: I
authors.lastname: Giles
authors.forename: David S
authors.initials: DS
authors.lastname: Latchman
authors.forename: David A
authors.initials: DA
authors.lastname: Isenberg
authors.forename: Anisur
authors.initials: A
authors.lastname: Rahman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001120:Arginine / Q000235:genetics* / Q000378:metabolism; D001324:Autoantigens; D001666:Binding Sites, Antibody / Q000235:genetics / Q000276:immunology; D019556:COS Cells; D004247:DNA / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D006657:Histones / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins / Q000276:immunology*; D017411:Ribonucleoproteins, Small Nuclear / Q000276:immunology*; D055517:snRNP Core Proteins; D000099207:SS-A Antigen
chemical_list: D000911:Antibodies, Monoclonal; D001120:Arginine; D001324:Autoantigens; D004247:DNA; D006657:Histones; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D055517:snRNP Core Proteins; D000099207:SS-A Antigen; C116006:RO60 protein, human