Crystallographic Analysis of 1,2,3–Trichloropropane
Biodegradation by Haloalkane Dehalogenase DhaA31

J 2014

Crystallographic Analysis of 1,2,3–Trichloropropane Biodegradation by Haloalkane Dehalogenase DhaA31

LAHODA, M., J.R. MESTERS, A. STSIAPANAVA, Radka CHALOUPKOVÁ, M. KUTY et. al.

Basic information

Original name

Crystallographic Analysis of 1,2,3–Trichloropropane Biodegradation by Haloalkane Dehalogenase DhaA31

Authors

LAHODA, M. (112 Belarus), J.R. MESTERS (724 Spain), A. STSIAPANAVA (112 Belarus), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), M. KUTY (203 Czech Republic), Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution) and I. KUTA SMATANOVA (203 Czech Republic)

Edition

Acta Crystallographica D, 2014, 0907-4449

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 7.232 in 2013

RIV identification code

RIV/00216224:14310/14:00074213

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1107/S1399004713026254

UT WoS

000331554500001

Keywords in English

1;2;3-trichloropropane; biodegradation; haloalkane dehalogenase DhaA31

Abstract

V originále

Haloalkane dehalogenases catalyze the hydrolytic cleavage of carbon-halogen bonds, which is a key step in aerobic mineralization of many environmental pollutants. One important pollutant is the toxic and anthropogenic compound 1,2,3-trichloropropane (TCP). Rational design was combined with saturation mutagenesis to obtain the haloalkane dehalogenase variant DhaA31, which displays an increased catalytic activity towards TCP. Here we report the 1.31 A crystal structure of substrate-free DhaA31, the 1.26 A structure of DhaA31 in a complex with TCP, and the 1.85 A wild-type DhaA structure. Crystals of the enzyme-substrate complex were successfully obtained by adding volatile TCP to the reservoir at crystallization at room temperature and pH 6.4. Comparison of the substrate-free structure with the DhaA31 enzyme-substrate complex reveals that the nucleophilic Asp106 changes its conformation from an inactive to an active state during the catalytic cycle. The positions of three chloride ions found inside the enzyme’s active site indicate a possible pathway for halide release from the active site through the main tunnel. Comparison of the mutant DhaA31 with wild-type DhaA revealed that introduced substitutions reduced volume and solvent accessibility of the active site pocket.

Links

GAP207/12/0775, research and development project

Name: Strukturně-funkční vztahy haloalkan dehalogenas

Investor: Czech Science Foundation

Citovat

LAHODA, M., J.R. MESTERS, A. STSIAPANAVA, Radka CHALOUPKOVÁ, M. KUTY, Jiří DAMBORSKÝ and I. KUTA SMATANOVA. Crystallographic Analysis of 1,2,3–Trichloropropane Biodegradation by Haloalkane Dehalogenase DhaA31. Acta Crystallographica D. 2014, vol. 70, february, p. 209-217. ISSN 0907-4449. Available from: https://dx.doi.org/10.1107/S1399004713026254.

@article{1213473,
   author = {Lahoda, M. and Mesters, J.R. and Stsiapanava, A. and Chaloupková, Radka and Kuty, M. and Damborský, Jiří and Kuta Smatanova, I.},
   article_number = {february},
   doi = {http://dx.doi.org/10.1107/S1399004713026254},
   keywords = {1;2;3-trichloropropane; biodegradation; haloalkane dehalogenase DhaA31},
   language = {eng},
   issn = {0907-4449},
   journal = {Acta Crystallographica D},
   title = {Crystallographic Analysis of 1,2,3–Trichloropropane Biodegradation by Haloalkane Dehalogenase DhaA31},
   volume = {70},
   year = {2014}
}

TY  - JOUR
ID  - 1213473
AU  - Lahoda, M. - Mesters, J.R. - Stsiapanava, A. - Chaloupková, Radka - Kuty, M. - Damborský, Jiří - Kuta Smatanova, I.
PY  - 2014
TI  - Crystallographic Analysis of 1,2,3–Trichloropropane Biodegradation by Haloalkane Dehalogenase DhaA31
JF  - Acta Crystallographica D
VL  - 70
IS  - february
SP  - 209-217
EP  - 209-217
SN  - 09074449
KW  - 1;2;3-trichloropropane
KW  - biodegradation
KW  - haloalkane dehalogenase DhaA31
N2  - Haloalkane dehalogenases catalyze the hydrolytic cleavage of carbon-halogen bonds, which is a key step in aerobic mineralization of many environmental pollutants. One important pollutant is the toxic and anthropogenic compound 1,2,3-trichloropropane (TCP). Rational design was combined with saturation mutagenesis to obtain the haloalkane dehalogenase variant DhaA31, which displays an increased catalytic activity towards TCP. Here we report the 1.31 A crystal structure of substrate-free DhaA31, the 1.26 A structure of DhaA31 in a complex with TCP, and the 1.85 A wild-type DhaA structure. Crystals of the enzyme-substrate complex were successfully obtained by adding volatile TCP to the reservoir at crystallization at room temperature and pH 6.4. Comparison of the substrate-free structure with the DhaA31 enzyme-substrate complex reveals that the nucleophilic Asp106 changes its conformation from an inactive to an active state during the catalytic cycle. The positions of three chloride ions found inside the enzyme’s active site indicate a possible pathway for halide release from the active site through the main tunnel. Comparison of the mutant DhaA31 with wild-type DhaA revealed that introduced substitutions reduced volume and solvent accessibility of the active site pocket.
ER  -

LAHODA, M., J.R. MESTERS, A. STSIAPANAVA, Radka CHALOUPKOVÁ, M. KUTY, Jiří DAMBORSKÝ and I. KUTA SMATANOVA. Crystallographic Analysis of 1,2,3–Trichloropropane Biodegradation by Haloalkane Dehalogenase DhaA31. \textit{Acta Crystallographica D}. 2014, vol.~70, february, p.~209-217. ISSN~0907-4449. Available from: https://dx.doi.org/10.1107/S1399004713026254.

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