| KOKKONEN, Piia Pauliina, J. SYKORA, Zbyněk PROKOP, A. GHOSE, David BEDNÁŘ, M. AMARO, Koen BEERENS, Šárka NEVOLOVÁ, Michaela SLÁNSKÁ, Jan BREZOVSKÝ, Jiří DAMBORSKÝ and M. HOF. Molecular Gating of an Engineered Enzyme Captured in Real Time. Journal of the American Chemical Society. Washington: American Chemical Society, 2018, vol. 140, No 51, p. 17999-18008. ISSN 0002-7863. Available from: https://dx.doi.org/10.1021/jacs.8b09848. |
Other formats:
BibTeX
LaTeX
RIS
@article{1503337,
author = {Kokkonen, Piia Pauliina and Sykora, J. and Prokop, Zbyněk and Ghose, A. and Bednář, David and Amaro, M. and Beerens, Koen and Nevolová, Šárka and Slánská, Michaela and Brezovský, Jan and Damborský, Jiří and Hof, M.},
article_location = {Washington},
article_number = {51},
doi = {http://dx.doi.org/10.1021/jacs.8b09848},
keywords = {PHOTOINDUCED ELECTRON-TRANSFER; CONFORMATIONAL DYNAMICS; CATALYTIC MECHANISM; PROTEIN DYNAMICS; NUCLEIC-ACIDS; ACTIVE-SITE; FORCE-FIELD; EVOLUTION; MOTIONS; SIMULATION},
language = {eng},
issn = {0002-7863},
journal = {Journal of the American Chemical Society},
title = {Molecular Gating of an Engineered Enzyme Captured in Real Time},
url = {https://loschmidt.chemi.muni.cz/peg/category/publications/#2018},
volume = {140},
year = {2018}
}
TY - JOUR
ID - 1503337
AU - Kokkonen, Piia Pauliina - Sykora, J. - Prokop, Zbyněk - Ghose, A. - Bednář, David - Amaro, M. - Beerens, Koen - Nevolová, Šárka - Slánská, Michaela - Brezovský, Jan - Damborský, Jiří - Hof, M.
PY - 2018
TI - Molecular Gating of an Engineered Enzyme Captured in Real Time
JF - Journal of the American Chemical Society
VL - 140
IS - 51
SP - 17999-18008
EP - 17999-18008
PB - American Chemical Society
SN - 00027863
KW - PHOTOINDUCED ELECTRON-TRANSFER
KW - CONFORMATIONAL DYNAMICS
KW - CATALYTIC MECHANISM
KW - PROTEIN DYNAMICS
KW - NUCLEIC-ACIDS
KW - ACTIVE-SITE
KW - FORCE-FIELD
KW - EVOLUTION
KW - MOTIONS
KW - SIMULATION
UR - https://loschmidt.chemi.muni.cz/peg/category/publications/#2018
N2 - Enzyme engineering tends to focus on the design of active sites for the chemical steps, while the physical steps of the catalytic cycle are often overlooked. Tight binding of a substrate in an active site is beneficial for the chemical steps, whereas good accessibility benefits substrate binding and product release. Many enzymes control the accessibility of their active sites by molecular gates. Here we analyzed the dynamics of a molecular gate artificially introduced into an access tunnel of the most efficient haloalkane dehalogenase using pre-steady-state kinetics, single-molecule fluorescence spectroscopy, and molecular dynamics. Photoinduced electron-transfer fluorescence correlation spectroscopy (PET-FCS) has enabled real-time observation of molecular gating at the single-molecule level with rate constants (k(on) = 1822 s(-1), k(off) = 60 s(-1)) corresponding well with those from the pre-steady-state kinetics (k(-1) = 1100 s(-1), k(1) = 20 s(-1)). The PET-FCS technique is used here to study the conformational dynamics in a soluble enzyme, thus demonstrating an additional application for this method. Engineering dynamical molecular gates represents a widely applicable strategy for designing efficient biocatalysts.
ER -
KOKKONEN, Piia Pauliina, J. SYKORA, Zbyněk PROKOP, A. GHOSE, David BEDNÁŘ, M. AMARO, Koen BEERENS, Šárka NEVOLOVÁ, Michaela SLÁNSKÁ, Jan BREZOVSKÝ, Jiří DAMBORSKÝ and M. HOF. Molecular Gating of an Engineered Enzyme Captured in Real Time. \textit{Journal of the American Chemical Society}. Washington: American Chemical Society, 2018, vol.~140, No~51, p.~17999-18008. ISSN~0002-7863. Available from: https://dx.doi.org/10.1021/jacs.8b09848.
|
