Real time measurements of elongation by a reverse transcriptase using surface plasmon resonance
| Autor: | Roy Williams, Malcolm Buckle, Henri Buc, Matteo Negroni |
|---|---|
| Přispěvatelé: | Physicochimie des Macromolecules Biologiques, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 1996 |
| Předmět: |
MESH: Streptavidin
Biosensing Techniques MESH: Base Sequence 01 natural sciences Surface plasmon resonance MESH: Bacterial Proteins Polymerase 0303 health sciences MESH: Dideoxynucleotides Multidisciplinary biology MESH: DNA Nucleic Acid Heteroduplexes MESH: Zidovudine RNA-Directed DNA Polymerase MESH: Thymine Nucleotides Reverse Transcriptase Inhibitors Elongation Zidovudine MESH: Biosensing Techniques Research Article Protein Binding Molecular Sequence Data Biotin MESH: Moloney murine leukemia virus 010402 general chemistry 03 medical and health sciences MESH: RNA-Directed DNA Polymerase Bacterial Proteins MESH: Nucleic Acid Heteroduplexes MESH: Biotin MESH: Protein Binding Thymine Nucleotides [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology 030304 developmental biology MESH: Molecular Sequence Data Base Sequence Processivity DNA Molecular biology Reverse transcriptase 0104 chemical sciences Polymerization Coding strand biology.protein Biophysics Streptavidin Primer (molecular biology) MESH: Reverse Transcriptase Inhibitors Moloney murine leukemia virus Dideoxynucleotides |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, 1996, 93 (2), pp.889-94. ⟨10.1073/pnas.93.2.889⟩ Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 1996, 93 (2), pp.889-94. ⟨10.1073/pnas.93.2.889⟩ |
| ISSN: | 0027-8424 1091-6490 |
| DOI: | 10.1073/pnas.93.2.889⟩ |
| Popis: | International audience; A rapid direct assay for polymerase-induced elongation along a given template is an obligate requirement for understanding the processivity of polymerization and the mode of action of drugs and inhibitors on this process. Surface plasmon resonance can be used to follow the association and the dissociation rates of a given reverse transcriptase on DNA.RNA and DNA.DNA hybrids immobilized on a biotin-streptavidin surface. The addition of nucleotides complementary to the template strand produces an increase in the local mass, as deduced from an increase in the measured signal, due to elongation of the primer strand that allows an estimation of both the extent and rate of the polymerization process. The terminator drug 3'-deoxy-3'-azidothymidine triphosphate completely abolishes the increase in signal as would be expected from an inhibition of elongation. This technique provides a sensitive assay for the affinities of different polymerases for specific templates and for the effects of terminators of the elongation process. |
| Databáze: | OpenAIRE |
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