Characterization of anti-Z-DNA antibody binding sites on Z-DNA by nuclear magnetic resonance spectroscopy

Two monoclonal anti-Z-DNA antibodies, Z22 and Z44, were shown to bind to the oligonucleotides, d(CG)2 and d(CG)3, and to interact with different parts of the helix. 1H nuclear magnetic resonance spectroscopy showed that Fab fragments stabilize an ordered structure in the tetranucleotide d(CG)2. Nucl...

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Bibliographic Details
Published in:The Journal of biological chemistry 1990-10, Vol.265 (30), p.18608-18614
Main Authors: Sanford, D G, Stollar, B D
Format: Article
Language:English
Subjects:
Animals
Antibodies, Antinuclear - metabolism
Antibodies, Monoclonal
Binding Sites, Antibody
Biological and medical sciences
DNA - immunology
Fundamental and applied biological sciences. Psychology
Immunoglobulin Fab Fragments - metabolism
In Vitro Techniques
Interactions. Associations
Intermolecular phenomena
Magnetic Resonance Spectroscopy
Mice
Models, Molecular
Molecular biophysics
Nucleic Acid Conformation
Oligodeoxyribonucleotides - metabolism
Protein Binding
Protein Conformation
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Summary:Two monoclonal anti-Z-DNA antibodies, Z22 and Z44, were shown to bind to the oligonucleotides, d(CG)2 and d(CG)3, and to interact with different parts of the helix. 1H nuclear magnetic resonance spectroscopy showed that Fab fragments stabilize an ordered structure in the tetranucleotide d(CG)2. Nuclear Overhauser effects measured in the presence of Z22 Fab indicate a syn conformation of guanine residues of d(CG)2. Intermolecular transfer of saturation between the Fabs and bound d(CG)3 was detected by a saturation of the protein spectrum and observation of changes in the DNA spectrum. Antibodies with deuterated aromatic amino acids were prepared to eliminate the protein aromatic resonances and thereby allow a more detailed analysis of the transfer to the DNA base protons. The greatest transfer with Z44 was to the dC-5 protons although all of the base protons interact with this antibody. Little, if any, transfer to the DNA base protons was observed with Z22. These results are consistent with a Z44 binding site on the convex surface of the Z-helix (analogous to the major groove of B-DNA) and a Z22 binding site on the sugar-phosphate backbone.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(17)44795-8