High resolution epitope mapping of an anti-DNA autoantibody using model DNA ligands

Patrick C. Swanson, Brian C. Cooper, Gary D. Glick

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

Although Ab-DNA complexes are involved in the pathogenesis of SLE, the mechanism by which anti-DNA recognize their DNA targets remains elusive. We have designed and synthesized small test DNA ligands that model ssDNA and dsDNA determinants commonly recognized by anti-DNA autoantibodies. These ligands were used in DNA footprinting assays to conduct a high resolution epitope-mapping study of anti-ssDNA BV04-01, which is typical of anti-ssDNA expressed in (NZB x NZW)F1 mice. We find that BV04-01 recognizes the ssDNA, but not dsDNA, epitopes presented on the test ligands, and makes specific base and backbone contacts with the DNA. Moreover, BV04-01 binding induces conformational changes in both the single-stranded and duplex region of the ligands. The epitope-mapping data is used in conjunction with computational molecular modeling to construct a three-dimensional model of a BV04-01- ligand complex. The salient aspects of this model are consistent with many features observed in the crystal structure of BV04-01 liganded to d(pT)3. This work extends previous anti-DNA binding studies and attempts to provide a unified approach that draws upon the merits of many precedented experimental methods to assemble three-dimensional structures of anti-DNA-DNA complexes. Analysis of these structures may ultimately help guide the development of molecules designed to inhibit anti-DNA binding in SLE.

Original languageEnglish (US)
Pages (from-to)2601-2612
Number of pages12
JournalJournal of Immunology
Volume152
Issue number5
StatePublished - Mar 1 1994
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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