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Specific Antibody-DNA Interaction: A Novel Strategy for Tight DNA Recognition

Published

Author(s)

S M. Di Pietro, J M. Centeno, M L. Cerutti, M F. Lodeiro, D U. Ferreiro, L G. Alonso, Frederick P. Schwarz, F Goldbaum, G de Prat-Gay

Abstract

Taking advantage of the extreme stability of a protein-DNA complex, we obtained highly specific monoclonal antibodies against a predetermined palindromic double stranded DNA sequence, corresponding to the binding site of the E2 transcriptional regulator of the human papillomavirus. The antibodies bind to the DNA solution with dissociation constants in the low and sub-nanomolar range, matching the affinity of the natural DNA binding domain. These antibodies discriminate effectively among a number of double and single stranded synthetic DNAs with factors ranging from 125 to 20,000-fold the dissociation constant of the specific DNA sequence used as the immunogen. We show that the recognition strategy of the antibodies is radically different from that of E2 and other natural DNA binders based on: 1) the resistance to high salt effects, 2) an entropically driven binding, 3) a postive heat capacity change of binding, and 4) spectroscopic evidence of base unstacking and partial denaturation of bound DNA.
Citation
Biochemistry
Volume
42
Issue
20

Keywords

antibody, biacore, calorimetry, dissociation constant, DNA

Citation

Di Pietro, S. , Centeno, J. , Cerutti, M. , Lodeiro, M. , Ferreiro, D. , Alonso, L. , Schwarz, F. , Goldbaum, F. and de Prat-Gay, G. (2003), Specific Antibody-DNA Interaction: A Novel Strategy for Tight DNA Recognition, Biochemistry (Accessed May 29, 2024)

Issues

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Created May 26, 2003, Updated October 12, 2021