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.

Základní údaje

Originální název

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

Autoři

LAHODA, M.; J.R. MESTERS; A. STSIAPANAVA; Radka CHALOUPKOVÁ; M. KUTY; Jiří DAMBORSKÝ a I. KUTA SMATANOVA

Vydání

Acta Crystallographica D, 2014, 0907-4449

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 7.232 v roce 2013

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/14:00074213

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

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

Štítky

AKR, rivok

Změněno: 28. 4. 2015 09:21, Ing. Andrea Mikešková

Anotace

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.

Návaznosti

GAP207/12/0775, projekt VaV

Název: Strukturně-funkční vztahy haloalkan dehalogenas

Investor: Grantová agentura ČR, Structure-functional Relationships of Haloalkane Dehalogenases

Citovat

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

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