Role of conformational dynamics in sequence-specific autoantibody*ssDNA recognition.
11F8 is a sequence-specific monoclonal anti-ssDNA autoantibody isolated from a lupus prone mouse that forms pathogenic complexes with ssDNA, resulting in kidney damage. Prior studies show that specificity is mediated by a somatic mutation from serine at (31)V(H) to arginine. Reversion back to serine in 11F8 resulted in >30-fold decrease in affinity and altered thermodynamic and kinetic parameters for sequence-specific recognition of its cognate ssDNA ligand. Mutagenesis and structural studies suggest that (R31)V(H) contacts ssDNA via a salt bridge and a bidentate hydrogen bond and may further contribute to specificity by altering binding-site conformation. Fluorescence resonance energy transfer experiments were conducted to assess the kinetics of conformational change during 11F8*ssDNA association. The extent of rearrangement between the six complementary determining regions in the 11F8*ssDNA complex with germline serine or somatically mutated arginine at residue 31 of the heavy chain was examined. Our studies show that greater conformational change occurs in five of six complementarity determining regions after the heavy chain germline J558 sequence undergoes mutation to arginine at (31)V(H).
Bobeck
Melissa J
MJ
Department of Chemistry, University of Michigan, Ann Arbor, MI 48109-1055, USA.
Glick
Gary D
GD
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D001323:Autoantibodies / Q000737:chemistry* / Q000276:immunology*; D001483:Base Sequence; D001666:Binding Sites, Antibody; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000276:immunology*; D031541:Fluorescence Resonance Energy Transfer; D006860:Hydrogen Bonding; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D013816:Thermodynamics; D013997:Time Factors
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D004277:DNA, Single-Stranded