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

J 2019

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

PRUDNIKOVA, T., B. KASCAKOVA, J.R. MESTERS, P. GRINKEVICH, P. HAVLICKOVA et. al.

Basic information

Original name

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

Authors

PRUDNIKOVA, T., B. KASCAKOVA, J.R. MESTERS, P. GRINKEVICH, P. HAVLICKOVA, A. MAZUR, A. SHAPOSHNIKOVA, Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution), M. KUTY and I.K. SMATANOVA

Edition

CRYSTALS, BASEL, MDPI, 2019, 2073-4352

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

20500 2.5 Materials engineering

Country of publisher

Switzerland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.404

RIV identification code

RIV/00216224:14310/19:00108205

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.3390/cryst9070375

UT WoS

000482052800024

Keywords in English

Haloalkane dehalogenase; halide-binding site; random microseeding

Abstract

V originále

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 Delta 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 Delta Cl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of the DbeA Delta Cl has been determined and refined to the 1.4 angstrom resolution. The DbeA Delta Cl crystals belong to monoclinic space group C121. The DbeA Delta Cl molecular structure was characterized and compared with five known haloalkane dehalogenases selected from the Protein Data Bank.

Links

ED0001/01/01, research and development project

Name: CETOCOEN

GA17-24321S, research and development project

Name: Studium hydratace a flexibility enzymů pomocí pokročilých strukturních a biofyzikálních metod

Investor: Czech Science Foundation

LM2015055, research and development project

Name: Centrum pro systémovou biologii (Acronym: C4SYS)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

PRUDNIKOVA, T., B. KASCAKOVA, J.R. MESTERS, P. GRINKEVICH, P. HAVLICKOVA, A. MAZUR, A. SHAPOSHNIKOVA, Radka CHALOUPKOVÁ, Jiří DAMBORSKÝ, M. KUTY and I.K. SMATANOVA. Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site. CRYSTALS. BASEL: MDPI, 2019, vol. 9, No 7, p. 1-12. ISSN 2073-4352. Available from: https://dx.doi.org/10.3390/cryst9070375.

@article{1639538,
   author = {Prudnikova, T. and Kascakova, B. and Mesters, J.R. and Grinkevich, P. and Havlickova, P. and Mazur, A. and Shaposhnikova, A. and Chaloupková, Radka and Damborský, Jiří and Kuty, M. and Smatanova, I.K.},
   article_location = {BASEL},
   article_number = {7},
   doi = {http://dx.doi.org/10.3390/cryst9070375},
   keywords = {Haloalkane dehalogenase; halide-binding site; random microseeding},
   language = {eng},
   issn = {2073-4352},
   journal = {CRYSTALS},
   title = {Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site},
   url = {http://dx.doi.org/10.3390/cryst9070375},
   volume = {9},
   year = {2019}
}

TY  - JOUR
ID  - 1639538
AU  - Prudnikova, T. - Kascakova, B. - Mesters, J.R. - Grinkevich, P. - Havlickova, P. - Mazur, A. - Shaposhnikova, A. - Chaloupková, Radka - Damborský, Jiří - Kuty, M. - Smatanova, I.K.
PY  - 2019
TI  - Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site
JF  - CRYSTALS
VL  - 9
IS  - 7
SP  - 1-12
EP  - 1-12
PB  - MDPI
SN  - 20734352
KW  - Haloalkane dehalogenase
KW  - halide-binding site
KW  - random microseeding
UR  - http://dx.doi.org/10.3390/cryst9070375
L2  - http://dx.doi.org/10.3390/cryst9070375
N2  - 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 Delta 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 Delta Cl was crystallized using the sitting-drop vapour-diffusion procedure with further optimization by the random microseeding technique. The crystal structure of the DbeA Delta Cl has been determined and refined to the 1.4 angstrom resolution. The DbeA Delta Cl crystals belong to monoclinic space group C121. The DbeA Delta Cl molecular structure was characterized and compared with five known haloalkane dehalogenases selected from the Protein Data Bank.
ER  -

PRUDNIKOVA, T., B. KASCAKOVA, J.R. MESTERS, P. GRINKEVICH, P. HAVLICKOVA, A. MAZUR, A. SHAPOSHNIKOVA, Radka CHALOUPKOVÁ, Jiří DAMBORSKÝ, M. KUTY and I.K. SMATANOVA. Crystallization and Crystallographic Analysis of a Bradyrhizobium Elkanii USDA94 Haloalkane Dehalogenase Variant with an Eliminated Halide-Binding Site. \textit{CRYSTALS}. BASEL: MDPI, 2019, vol.~9, No~7, p.~1-12. ISSN~2073-4352. Available from: https://dx.doi.org/10.3390/cryst9070375.

Detailed Information on Publication Record