Crystallization and Crystallographic Characterization of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site

Authors

Prudnikova, T., Kascakova, B., Mesters, J. R., Grinkevich, P., Havlickova, P., Mazur, A., Shaposhnikova, A., Chaloupkova, R., Damborsky, J., Kuty, M., Kuta Smatanova, I.

Source

CRYSTALS 9: 375 (2019)

Abstract

Haloalkane dehalogenases are a very important class of microbial enzymes for environmental detoxification of halogenated pollutants, for biocatalysis, biosensing and molecular tagging. The double mutant (Ile44Leu + Gln102His) of the haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 (DbeA∆Cl) was constructed to study the role of the second halide-binding site previously discovered in the wild-type structure. The variant is less active, less stable in the presence of chloride ions and exhibits significantly altered substrate specificity when compared with the DbeAwt. DbeA∆Cl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of
the DbeA∆Cl has been determined and refined to the 1.4 Å resolution. The DbeA∆Cl crystals belong to monoclinic space group C121. The DbeA∆Cl molecular structure was characterized and compared
with five known haloalkane dehalogenases selected from the Protein Data Bank.

Full text

Citation

Prudnikova, T., Kascakova, B., Mesters, J. R., Grinkevich, P., Havlickova, P., Mazur, A., Shaposhnikova, A., Chaloupkova, R., Damborsky, J., Kuty, M., Kuta Smatanova, I., 2019: Crystallization and Crystallographic Characterization of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site. Crystals 9: 375.

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