Artículo
Cerutti, M.L.; Centeno, J.M.; Goldbaum, F.A.; De Prat-Gay, G. "Generation of Sequence-specific, High Affinity Anti-DNA Antibodies" (2001) Journal of Biological Chemistry. 276(16):12769-12773
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Abstract:
By taking advantage of the extreme stability of a protein-DNA complex, we have obtained two highly specific monoclonal antibodies against a predetermined palindromic DNA sequence corresponding to the binding site of the E2 transcriptional regulator of the human papillomavirus (HPV-16). The purified univalent antibody fragments bind to a double-stranded DNA oligonucleotide corresponding to the E2 binding site in solution with dissociation constants in the low and subnanomolar range. This affinity matches that of the natural DNA binding domain and is severalfold higher than the affinity of a homologous bovine E2 C-terminal domain (BPV-1) for the same DNA. 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 in the immunogenic protein-DNA complex. Moreover, they are capable of fine specificity tuning, since they both bind less tightly to another HPV-16 E2 binding site, differing in only 1 base pair in a noncontact flexible region. Beyond the relevance of obtaining a specific anti-DNA response, these results provide a first glance at how DNA as an antigen is recognized specifically by an antibody. The accuracy of the spectroscopic method used for the binding analysis suggests that a detailed mechanistic analysis is attainable.
Registro:
| Documento: | Artículo |
| Título: | Generation of Sequence-specific, High Affinity Anti-DNA Antibodies |
| Autor: | Cerutti, M.L.; Centeno, J.M.; Goldbaum, F.A.; De Prat-Gay, G. |
| Filiación: | Inst. de Investigaciones Bioquimicas, Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Patricias Argentinas 435, (1405) Buenos Aires, Argentina |
| Palabras clave: | Antigens; Chemical bonds; Complexation; Dissociation; DNA; DNA sequences; Spectroscopy; Dissociation constants; Antibodies; DNA; DNA antibody; oligonucleotide; transcription factor E2; unclassified drug; DNA; DNA binding protein; E2 protein, Bovine papillomavirus; E2 protein, Human papillomavirus type 16; monoclonal antibody; oncoprotein; virus protein; antibody specificity; antigen binding; antigen recognition; article; binding affinity; binding assay; binding site; carboxy terminal sequence; complex formation; dissociation constant; DNA binding; DNA sequence; immunogenicity; priority journal; protein DNA binding; protein DNA interaction; spectroscopy; transcription regulation; amino acid sequence; animal; antibody combining site; cattle; chemistry; enzyme linked immunosorbent assay; human; immunoglobulin variable region; immunology; metabolism; molecular genetics; mouse; Papilloma virus; sequence alignment; sequence homology; Bovinae; Human papillomavirus; Human papillomavirus type 16; Papillomavirus; Amino Acid Sequence; Animals; Antibodies, Monoclonal; Binding Sites; Binding Sites, Antibody; Cattle; DNA; DNA-Binding Proteins; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulin Variable Region; Mice; Molecular Sequence Data; Oncogene Proteins, Viral; Papillomaviridae; Sequence Alignment; Sequence Homology, Amino Acid; Viral Proteins |
| Año: | 2001 |
| Volumen: | 276 |
| Número: | 16 |
| Página de inicio: | 12769 |
| Página de fin: | 12773 |
| DOI: | http://dx.doi.org/10.1074/jbc.M100260200 |
| Título revista: | Journal of Biological Chemistry |
| Título revista abreviado: | J. Biol. Chem. |
| ISSN: | 00219258 |
| CODEN: | JBCHA |
| CAS: | DNA, 9007-49-2; Antibodies, Monoclonal; Binding Sites, Antibody; DNA, 9007-49-2; DNA-Binding Proteins; E2 protein, Bovine papillomavirus; E2 protein, Human papillomavirus type 16; Immunoglobulin Variable Region; Oncogene Proteins, Viral; Viral Proteins |
| PDF: | https://bibliotecadigital.exactas.uba.ar/download/paper/paper_00219258_v276_n16_p12769_Cerutti.pdf |
| Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v276_n16_p12769_Cerutti |
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Citas:
---------- APA ----------
Cerutti, M.L., Centeno, J.M., Goldbaum, F.A. & De Prat-Gay, G. (2001) . Generation of Sequence-specific, High Affinity Anti-DNA Antibodies. Journal of Biological Chemistry, 276(16), 12769-12773.http://dx.doi.org/10.1074/jbc.M100260200
---------- CHICAGO ----------
Cerutti, M.L., Centeno, J.M., Goldbaum, F.A., De Prat-Gay, G. "Generation of Sequence-specific, High Affinity Anti-DNA Antibodies" . Journal of Biological Chemistry 276, no. 16 (2001) : 12769-12773.http://dx.doi.org/10.1074/jbc.M100260200
---------- MLA ----------
Cerutti, M.L., Centeno, J.M., Goldbaum, F.A., De Prat-Gay, G. "Generation of Sequence-specific, High Affinity Anti-DNA Antibodies" . Journal of Biological Chemistry, vol. 276, no. 16, 2001, pp. 12769-12773.http://dx.doi.org/10.1074/jbc.M100260200
---------- VANCOUVER ----------
Cerutti, M.L., Centeno, J.M., Goldbaum, F.A., De Prat-Gay, G. Generation of Sequence-specific, High Affinity Anti-DNA Antibodies. J. Biol. Chem. 2001;276(16):12769-12773.http://dx.doi.org/10.1074/jbc.M100260200
