Thermodynamic basis for sequence-specific recognition of ssDNA by an autoantibody.
11F8 is a sequence-specific DNA binding monoclonal autoantibody previously isolated from an autoimmune lupus-prone mouse [Stevens, S. Y., and Glick, G. D. (1999) Biochemistry 38, 560-568]. This antibody, like many other lupus anti-DNAs, localizes to kidney tissue and eventually leads to renal damage through a process that first involves the binding of DNA antigens. A series of experiments were conducted to investigate the thermodynamic and structural basis by which this antibody discriminates between specific, noncognate, and nonspecific sequences. Sequence-specific binding occurs with a minimal dependence on the polyelectrolyte effect along with a favorable binding enthalpy reflecting the presence of base stacking and contacts to DNA bases. This favorable binding enthalpy apparently is derived from desolvation at the binding interface and is consistent with recent models of the nonclassical hydrophobic effect. Noncognate recognition is also driven by the nonclassical hydrophobic effect, but is accompanied by highly unfavorable entropies that are responsible for reduced affinity relative to the high-affinity consensus sequence. Nonspecific recognition is driven completely by the polyelectrolyte effect involving extensive electrostatic interactions with the phosphate backbone. Collectively, the data demonstrate the ability of 11F8 to adapt its mode of binding to the available DNA surface and provide a thermodynamic model for sequence-specific recognition of single-stranded DNA. The salient features of this model employ the paradigms invoked to explain protein.dsDNA, protein.RNA, and antibody.antigen binding.
Departments of Chemistry and Biological Chemistry, The University of Michigan, 930 North University Avenue, Ann Arbor, Michigan 48109-1055, USA.
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Ackroyd
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Cleary
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Glick
D000818:Animals; D000974:Antibodies, Antinuclear / Q000737:chemistry / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D000915:Antibody Affinity; D001483:Base Sequence; D001666:Binding Sites, Antibody; D018983:DNA Footprinting; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D019277:Entropy; D006860:Hydrogen Bonding; D051379:Mice; D009690:Nucleic Acid Conformation; D011196:Potassium Permanganate / Q000737:chemistry; D012492:Salts / Q000737:chemistry; D019346:Sodium Acetate / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D012997:Solvents; D013696:Temperature; D013816:Thermodynamics
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D012492:Salts; D012997:Solvents; D011196:Potassium Permanganate; D012965:Sodium Chloride; D019346:Sodium Acetate