VALUCHOVÁ, Soňa, Jaroslav FULNEČEK, Alexander PETROV, Konstantinos TRIPSIANES and Karel ŘÍHA. A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement. Scientific Reports. LONDON: NATURE PUBLISHING GROUP, 2016, vol. 6, December, p. nestránkováno, 10 pp. ISSN 2045-2322. Available from: https://dx.doi.org/10.1038/srep39653. |
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@article{1375066, author = {Valuchová, Soňa and Fulneček, Jaroslav and Petrov, Alexander and Tripsianes, Konstantinos and Říha, Karel}, article_location = {LONDON}, article_number = {December}, doi = {http://dx.doi.org/10.1038/srep39653}, keywords = {SINGLE-MOLECULE; ENDONUCLEASE XPF-ERCC1; TRANSCRIPTION FACTORS; DNA INTERACTIONS; BINDING; REPAIR; POLYMERASE; INHIBITORS; ANISOTROPY; MECHANISM}, language = {eng}, issn = {2045-2322}, journal = {Scientific Reports}, title = {A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement}, url = {http://www.nature.com/articles/srep39653}, volume = {6}, year = {2016} }
TY - JOUR ID - 1375066 AU - Valuchová, Soňa - Fulneček, Jaroslav - Petrov, Alexander - Tripsianes, Konstantinos - Říha, Karel PY - 2016 TI - A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement JF - Scientific Reports VL - 6 IS - December SP - nestránkováno EP - nestránkováno PB - NATURE PUBLISHING GROUP SN - 20452322 KW - SINGLE-MOLECULE KW - ENDONUCLEASE XPF-ERCC1 KW - TRANSCRIPTION FACTORS KW - DNA INTERACTIONS KW - BINDING KW - REPAIR KW - POLYMERASE KW - INHIBITORS KW - ANISOTROPY KW - MECHANISM UR - http://www.nature.com/articles/srep39653 L2 - http://www.nature.com/articles/srep39653 N2 - Many fundamental biological processes depend on intricate networks of interactions between proteins and nucleic acids and a quantitative description of these interactions is important for understanding cellular mechanisms governing DNA replication, transcription, or translation. Here we present a versatile method for rapid and quantitative assessment of protein/nucleic acid (NA) interactions. This method is based on protein induced fluorescence enhancement (PIFE), a phenomenon whereby protein binding increases the fluorescence of Cy3-like dyes. PIFE has mainly been used in single molecule studies to detect protein association with DNA or RNA. Here we applied PIFE for steady state quantification of protein/NA interactions by using microwell plate fluorescence readers (mwPIFE). We demonstrate the general applicability of mwPIFE for examining various aspects of protein/DNA interactions with examples from the restriction enzyme BamHI, and the DNA repair complexes Ku and XPF/ERCC1. These include determination of sequence and structure binding specificities, dissociation constants, detection of weak interactions, and the ability of a protein to translocate along DNA. mwPIFE represents an easy and high throughput method that does not require protein labeling and can be applied to a wide range of applications involving protein/NA interactions. ER -
VALUCHOVÁ, Soňa, Jaroslav FULNEČEK, Alexander PETROV, Konstantinos TRIPSIANES and Karel ŘÍHA. A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement. \textit{Scientific Reports}. LONDON: NATURE PUBLISHING GROUP, 2016, vol.~6, December, p.~nestránkováno, 10 pp. ISSN~2045-2322. Available from: https://dx.doi.org/10.1038/srep39653.
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