Effect of antibody excess on the size, stoichiometry, and DNAse resistance of DNA anti-DNA immune complexes.
The binding of antibodies to DNA was examined under conditions of increasing antibody excess. DNA anti-DNA immune complexes (IC) formed at increasing antibody to DNA ratios were digested with excess DNAse I, and the DNAse-resistant (protected) IC were analyzed. With increasing antibody excess, the size of the IC that were resistant to DNAse digestion increased, and the size of the protected DNA within the IC also increased. This suggested that IgG molecules could bind in close proximity along the DNA molecule, preventing access of DNAse to the DNA between adjacent IgG. To further define the binding of adjacent IgG, DNAse digested IC containing one or two IgG were isolated, and the DNA contained within these IC was analyzed on DNA sequencing gels. Binding of a single IgG to DNA resulted in the protection of a DNA fragment 35 to 45 base pairs (bp) long, corresponding to the distance between binding sites of a single IgG molecule. Binding of two IgG to DNA protected a DNA fragment 50 to 60 bp long, 1 1/2 times the size of the fragment protected by one IgG. These data suggest that in conditions of Ab excess, IgG molecules can interdigitate along the DNA molecule, resulting in small, stable, DNAse-resistant IC of high antibody density.
D000974:Antibodies, Antinuclear / Q000502:physiology*; D000936:Antigen-Antibody Complex; D001482:Base Composition; D001666:Binding Sites, Antibody; D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D003851:Deoxyribonucleases; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D046911:Macromolecular Substances; D008433:Mathematics
D000974:Antibodies, Antinuclear; D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D004247:DNA; D003851:Deoxyribonucleases