Nanosecond Time-Dependent Stokes Shift at the Tunnel Mouth of Haloalkane Dehalogenases

Jesenská, A.; Sýkora, Jan; Olžyńska, Agnieszka; Brezovský, J.; Zdráhal, Z.; Damborský, J.; Hof, Martin

doi:https://dx.doi.org/10.1021/ja804020q

Number of the records: 1

Nanosecond Time-Dependent Stokes Shift at the Tunnel Mouth of Haloalkane Dehalogenases

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SYSNO ASEP	0333807
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Nanosecond Time-Dependent Stokes Shift at the Tunnel Mouth of Haloalkane Dehalogenases
Title	Časově závislý Stokesův posuv v blízkosti aktivního místa Haloalkan- Dehalogenaz
Author(s)	Jesenská, A. (CZ) Sýkora, Jan (UFCH-W)_RID Olžyńska, Agnieszka (UFCH-W)_RID Brezovský, J. (CZ) Zdráhal, Z. (CZ) Damborský, J. (CZ) Hof, Martin (UFCH-W)_{RID, ORCID}
Source Title	Journal of the American Chemical Society. - : American Chemical Society - ISSN 0002-7863 Roč. 131, č. 2 (2009), s. 494-501
Number of pages	8 s.
Language	eng - English
Country	US - United States
Keywords	crystallographic analysis ; active-site dynamics ; solvent relaxation
Subject RIV	CF - Physical ; Theoretical Chemistry
R&D Projects	GA203/08/0114 GA ČR - Czech Science Foundation (CSF)
	LC06063 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	LC06010 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
CEZ	AV0Z40400503 - UFCH-W (2005-2011)
UT WOS	000262521800036
DOI	10.1021/ja804020q
Annotation	The tunnel mouths are evolutionally the most variable regions in the structures of haloalkane dehalogenases originating from different bacterial species, suggesting their importance for adaptation of enzymes to various substrates. We decided to monitor the dynamics of this particular region by means of time-resolved fluorescence spectroscopy and molecular dynamic simulations. To label the enzyme specifically, we adapted a novel procedure that utilizes a coumarin dye containing a halide-hydrocarbon linker, which serves as a substrate for enzymatic reaction. The procedure leads to a coumarin dye covalently attached and specifically located in the tunnel mouth of the enzyme. In this manner, we stained two haloalkane dehalogenase mutants, DbjA-H280F and DhaA-H272F.
Workplace	J. Heyrovsky Institute of Physical Chemistry
Contact	Michaela Knapová, michaela.knapova@jh-inst.cas.cz, Tel.: 266 053 196
Year of Publishing	2010

Number of the records: 1