Detailed Information on Publication Record
2013
Are Time-Dependent Fluorescence Shifts at the Tunnel Mouth of Haloalkane Dehalogenase Enzymes Dependent on the Choice of the Chromophore?
AMARO, Mariana, Jan BREZOVSKÝ, Silvia KOVÁČOVÁ, Lukáš MAIER, Radka CHALOUPKOVÁ et. al.Basic information
Original name
Are Time-Dependent Fluorescence Shifts at the Tunnel Mouth of Haloalkane Dehalogenase Enzymes Dependent on the Choice of the Chromophore?
Authors
AMARO, Mariana (620 Portugal), Jan BREZOVSKÝ (203 Czech Republic, belonging to the institution), Silvia KOVÁČOVÁ (703 Slovakia, belonging to the institution), Lukáš MAIER (203 Czech Republic, belonging to the institution), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), Jan SYKORA (203 Czech Republic), Kamil PARUCH (203 Czech Republic, guarantor, belonging to the institution), Jiří DAMBORSKÝ (203 Czech Republic, belonging to the institution) and Martin HOF (203 Czech Republic)
Edition
Journal of Physical Chemistry B, WASHINGTON, AMER CHEMICAL SOC, 2013, 1520-6106
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
Genetics and molecular biology
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
Impact factor
Impact factor: 3.377
RIV identification code
RIV/00216224:14310/13:00065798
Organization unit
Faculty of Science
UT WoS
000321542200006
Keywords in English
DYNAMIC STOKES SHIFT; WATER-PROTEIN FLUCTUATIONS; POLAR SOLVATION DYNAMICS; HYDRATION DYNAMICS; DIELECTRIC RESPONSE; SOLVENT RELAXATION; ACTIVE-SITE; SUBSTRATE; SURFACE; SIMULATIONS
Změněno: 14/8/2013 08:45, Ing. Andrea Mikešková
Abstract
V originále
AB Time-dependent fluorescence shifts (TDFS) of chromophores selectively attached to proteins may give information on the dynamics of the probed protein moieties and their degree of hydration. Previously, we demonstrated that a coumarin dye selectively labeling the tunnel mouth of different haloalkane dehalogenases (HLDs) can distinguish between different widths of tunnel mouth openings. In order to generalize those findings analogous experiments were performed using a different chromophore probing the same region of these enzymes. To this end we synthesized and characterized three new fluorescent probes derived from dimethylaminonaphthalene bearing a linker almost identical to that of the coumarin dye used in our previous study. Labeling efficiencies, acrylamide quenching, fluorescence anisotropies, and TDFS for the examined fluorescent substrates confirm the picture gained from the coumarin studies: the different tunnel mouth opening, predicted by crystal structures, is reflected in the hydration and tunnel mouth dynamics of the investigated HLDs. Comparison of the TDFS reported by the coumarin dye with those obtained with the new dimethylaminonaphthalene dyes shows that the choice of chromophore may strongly influence the recorded TDFS characteristics. The intrinsic design of our labeling strategy and the variation of the linker length ensure that both dyes probe the identical enzyme region; moreover, the covalently fixed position of the chromophore does not allow for a major relocalization within the HLD structures. Our study shows, for the first time, that TDFS may strongly depend on the choice of the chromophore, even though the identical region of a protein is explored.
Links
CZ.1.05/2.1.00/01.0001, interní kód MU |
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GAP207/12/0775, research and development project |
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IAA401630901, research and development project |
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