| VALUCHOVÁ, Soňa, Jaroslav FULNEČEK, Alexander PETROV, Konstantinos TRIPSIANES a Karel ŘÍHA. A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement. Scientific Reports. LONDON: NATURE PUBLISHING GROUP, 2016, roč. 6, December, s. nestránkováno, 10 s. ISSN 2045-2322. Dostupné z: 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 a Karel ŘÍHA. A rapid method for detecting protein-nucleic acid interactions by protein induced fluorescence enhancement. \textit{Scientific Reports}. LONDON: NATURE PUBLISHING GROUP, 2016, roč.~6, December, s.~nestránkováno, 10 s. ISSN~2045-2322. Dostupné z: https://dx.doi.org/10.1038/srep39653.
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