The importance of somatic mutations in the V(lambda) gene 2a2 in human monoclonal anti-DNA antibodies.
2a2 is the most commonly rearranged gene in the human V(lambda )locus. It has been postulated that certain immunoglobulin genes (including 2a2) are rearranged preferentially because their germline sequences encode structures capable of binding to a range of antigens. Somatic mutation could then increase the specificity and affinity of binding to a particular antigen. We studied the properties of five IgG molecules in which the same heavy chain was paired with different light chains derived from 2a2. The pattern of somatic mutations in 2a2 was shown to be crucial in conferring the ability to bind DNA, but two different patterns of mutation each conferred this ability.Computer-generated models of the three-dimensional structures of these antibodies illustrate the ability of 2a2 to form a DNA binding site in different ways. Somatic mutations at the periphery of the DNA binding site were particularly important. In two different light chains, mutations to arginine at different sites in the complementarity determining regions (CDRs) enhanced binding to DNA. In a third light chain, however, mutation to arginine at a different site blocked binding to DNA.
Center for Rheumatology/Bloomsbury Rheumatology Unit, Division of Medicine, University College, London, UK. anisur.rahman@ucl.ac.uk
A
A
Rahman
J
J
Haley
E
E
Radway-Bright
S
S
Nagl
D G
DG
Low
D S
DS
Latchman
D A
DA
Isenberg
D000595:Amino Acid Sequence; D000974:Antibodies, Antinuclear / Q000737:chemistry / Q000235:genetics*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics*; D003198:Computer Simulation; D004247:DNA / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D004247:DNA