Crystallization and preliminary X-ray diffraction analysis of the wild-type haloalkane dehalogenase DhaA and its variant DhaA13 complexed with different ligands

0373764 - ÚMG 2012 RIV GB eng J - Journal Article
Stsiapanava, A. - Chaloupková, R. - Fořtová, A. - Brynda, Jiří - Weiss, M.S. - Damborský, J. - Kutá-Smatanová, Ivana
Crystallization and preliminary X-ray diffraction analysis of the wild-type haloalkane dehalogenase DhaA and its variant DhaA13 complexed with different ligands.
Acta Crystallographica Section F-Structural Biology and Crystallization Communications. Roč. 67, - (2011), s. 253-257. ISSN 1744-3091. E-ISSN 2053-230X
R&D Projects: GA MŠMT(CZ) LC06010
Grant - others:GA ČR(CZ) GA310/09/1407
Program: GA
Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z60870520
Keywords : haloalkane dehalogenases * DhaA * Rhodococcus rhodochrous * microseeding * atomic resolution
Subject RIV: EB - Genetics ; Molecular Biology; CD - Macromolecular Chemistry (UEK-B)
Impact factor: 0.506, year: 2011

Haloalkane dehalogenases make up an important class of hydrolytic enzymes which catalyse the cleavage of carbon-halogen bonds in halogenated aliphatic compounds. There is growing interest in these enzymes owing to their potential use in environmental and industrial applications. The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 can slowly detoxify the industrial pollutant 1,2,3-trichloropropane (TCP). Structural analysis of this enzyme complexed with target ligands was conducted in order to obtain detailed information about the structural limitations of its catalytic properties. In this study, the crystallization and preliminary X-ray analysis of complexes of wild-type DhaA with 2-propanol and with TCP and of complexes of the catalytically inactive variant DhaA13 with the dye coumarin and with TCP are described. The crystals of wild-type DhaA were plate-shaped and belonged to the triclinic space group P1, while the variant DhaA13 can form prism-shaped crystals belonging to the orthorhombic space group P2(1)2(1)2(1) as well as plate-shaped crystals belonging to the triclinic space group P1. Diffraction data for crystals of wild-type DhaA grown from crystallization solutions with different concentrations of 2-propanol were collected to 1.70 and 1.26 Å resolution, respectively. A prism-shaped crystal of DhaA13 complexed with TCP and a plate-shaped crystal of the same variant complexed with the dye coumarin diffracted X-rays to 1.60 and 1.33 Å resolution, respectively. A crystal of wild-type DhaA and a plate-shaped crystal of DhaA13, both complexed with TCP, diffracted to atomic resolutions of 1.04 and 0.97 Å, respectively.
Permanent Link: http://hdl.handle.net/11104/0206833