Location of Cyanine-3 on Double-Stranded DNA: Importance for Fluorescence Resonance Energy Transfer Studies
| Autor: | David M.J. Lilley, David Norman, Dušan Uhrín, Richard J. Grainger |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Models
Molecular Fluorophore Base Sequence Base pair fungi DNA Carbocyanines Biochemistry Fluorescence chemistry.chemical_compound Crystallography Spectrometry Fluorescence Förster resonance energy transfer Energy Transfer Oligodeoxyribonucleotides chemistry Helix Nucleic Acid Conformation Fluorescein Cyanine Nuclear Magnetic Resonance Biomolecular Fluorescent Dyes Macromolecule |
| Zdroj: | Biochemistry. 39:6317-6324 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi992944a |
| Popis: | Fluorescence resonance energy transfer provides valuable long-range distance information about macromolecules in solution. Fluorescein and Cy3 are an important donor-acceptor pair of fluorophores; the characteristic Förster length for this pair on DNA is 56 A, so the pair can be used to study relatively long distances. Measurement of FRET efficiency for a series of DNA duplexes terminally labeled with fluorescein and Cy3 suggests that the Cy3 is close to the helical axis of the DNA. An NMR analysis of a self-complementary DNA duplex 5'-labeled with Cy3 shows that the fluorophore is stacked onto the end of the helix, in a manner similar to that of an additional base pair. This provides a known point from which distances calculated from FRET measurements are measured. Using the FRET efficiencies for the series of DNA duplexes as restraints, we have determined an effective position for the fluorescein, which is maximally extended laterally from the helix. The knowledge of the fluorophore positions can now be used for more precise interpretation of FRET data from nucleic acids. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|