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 language | English |
|---|---|
| Pages (from-to) | 2601-2612 |
| Number of pages | 12 |
| Journal | Journal of Immunology |
| Volume | 152 |
| Issue number | 5 |
| State | Published - Mar 1 1994 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Immunology
Cite this
High resolution epitope mapping of an anti-DNA autoantibody using model DNA ligands. / Swanson, Patrick; Cooper, Brian C.; Glick, Gary D.
In: Journal of Immunology, Vol. 152, No. 5, 01.03.1994, p. 2601-2612.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - High resolution epitope mapping of an anti-DNA autoantibody using model DNA ligands
AU - Swanson, Patrick
AU - Cooper, Brian C.
AU - Glick, Gary D.
PY - 1994/3/1
Y1 - 1994/3/1
N2 - 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.
AB - 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.
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M3 - Article
VL - 152
SP - 2601
EP - 2612
JO - Journal of Immunology
JF - Journal of Immunology
SN - 0022-1767
IS - 5
ER -