DNA hydrolysis by monoclonal autoantibody BV 04-01 |
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| Authors: | L. S. Rodkey G. Gololobov C. A. Rumbley J. Rumbley D. V. Schourov O. I. Makarevich A. G. Gabibov E. W. Voss Jr |
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| Affiliation: | (1) Department of Pathology and Laboratory Medicine, University of Texas-Houston, Medical School, 77225-0708 Houston, TX;(2) Department of Cell and Structural Biology, University of Illinois at Urbana-Champaign, 505 South Goodwin Avenue, 61801, IL, Urbana;(3) Present address: Department of Medicine, Allergy and Immunology Section, University of Pennsylvania School of Medicine, 19104 Philadelphia, PA;(4) Department of Biochemistry, University of Illinois at Urbana-Champaign, 600 South Matthews, 61801 Urbana, IL;(5) Present address: Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, 19104 Philadelphia, PA;(6) Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilova 32, 117984 Moscow, Russia;(7) Department of Microbiology, University of Illinois at Urbana-Champaign, 407 South Goodwin Avenue, 61801 Urbana, IL;(8) Department of Pathology and Laboratory Medicine, University of Texas-Houston, Medical School, P.O. Box 20708, 77225-0708 Houston, TX |
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| Abstract: | Monoclonal anti-DNA autoantibody BV 04-01 catalyzed hydrolysis of DNA in the presence of Mg2+. Catalysis was asssociated with BV 04-01 IgG, Fab, and single-chain-antibody (SCA) proteins. Cleavage of both ss and dsDNA was observed with efficient hydrolysis of the C-rich region of A7C7ATATAGCGCGT2, as well as a preference for cleaving within CG-rich regions of dsDNA. Data nn specificity of ssDNA hydrolysis and kinetic data obtained from wild-type SCA, and two SCA mutants were used to model the catalytically active antibody site using the previously resolved X-ray structure of BV 04-01. The resulting model suggested that the target phosphodiester bond is activated by induction of conformational strain. In addition, the antibody-DNA complex contained a Mg2+ coordination site composed of the L32Tyr and L27d His side chains and a DNA 3′-phosphodiester group. Induction of strain along with the metal coordination could be part of the mechanism by which this antibody catalyzes DNA hydrolysis. Sequence data for BV 04-01 VH and VL genes suggested that the proposed catalytic-antibody active site was germline-encoded. This observation suggests that catalytic activity might represent an important—rarely examined—function for some antibody molecules. |
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| Keywords: | Catalysis autoantibody abzyme |
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