| CHALOUPKOVÁ, Radka, Veronika LIŠKOVÁ, Martin TOUL, Klára MARKOVÁ, Eva ŠEBESTOVÁ, Lenka HERNYCHOVÁ, Martin MAREK, José Gaspar RANGEL PAMPLONA PIZARRO PINTO, Daniel PLUSKAL, Jitka WATERMAN, Zbyněk PROKOP and Jiří DAMBORSKÝ. Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts. ACS Catalysis. Washington, D.C.: AMER CHEMICAL SOC, 2019, vol. 9, No 6, p. 4810-4823. ISSN 2155-5435. Available from: https://dx.doi.org/10.1021/acscatal.9b01031. |
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@article{1545949,
author = {Chaloupková, Radka and Lišková, Veronika and Toul, Martin and Marková, Klára and Šebestová, Eva and Hernychová, Lenka and Marek, Martin and Rangel Pamplona Pizarro Pinto, José Gaspar and Pluskal, Daniel and Waterman, Jitka and Prokop, Zbyněk and Damborský, Jiří},
article_location = {Washington, D.C.},
article_number = {6},
doi = {http://dx.doi.org/10.1021/acscatal.9b01031},
keywords = {catalytic promiscuity; ancestral reconstruction; haloalkane dehalogenase; monooxygenase; luciferase; emergence of biological function},
language = {eng},
issn = {2155-5435},
journal = {ACS Catalysis},
title = {Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts},
url = {https://pubs.acs.org/doi/abs/10.1021/acscatal.9b01031},
volume = {9},
year = {2019}
}
TY - JOUR
ID - 1545949
AU - Chaloupková, Radka - Lišková, Veronika - Toul, Martin - Marková, Klára - Šebestová, Eva - Hernychová, Lenka - Marek, Martin - Rangel Pamplona Pizarro Pinto, José Gaspar - Pluskal, Daniel - Waterman, Jitka - Prokop, Zbyněk - Damborský, Jiří
PY - 2019
TI - Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts
JF - ACS Catalysis
VL - 9
IS - 6
SP - 4810-4823
EP - 4810-4823
PB - AMER CHEMICAL SOC
SN - 21555435
KW - catalytic promiscuity
KW - ancestral reconstruction
KW - haloalkane dehalogenase
KW - monooxygenase
KW - luciferase
KW - emergence of biological function
UR - https://pubs.acs.org/doi/abs/10.1021/acscatal.9b01031
L2 - https://pubs.acs.org/doi/abs/10.1021/acscatal.9b01031
N2 - To obtain structural insights into the emergence of biological functions from catalytically promiscuous enzymes, we reconstructed an ancestor of catalytically distinct, but evolutionarily related, haloalkane dehalogenases (EC 3.8.1.5) and Renilla luciferase (EC 1.13.12.5). This ancestor has both hydrolase and monooxygenase activities. Its crystal structure solved to 1.39 angstrom resolution revealed the presence of a catalytic pentad conserved in both dehalogenase and luciferase descendants and a molecular oxygen bound in between two residues typically stabilizing a halogen anion. The differences in the conformational dynamics of the specificity-determining cap domains between the ancestral and descendant enzymes were accessed by molecular dynamics and hydrogen-deuterium exchange mass spectrometry. Stopped-flow analysis revealed that the alkyl enzyme intermediate formed in the luciferase-catalyzed reaction is trapped by blockage of a hydrolytic reaction step. A single-point mutation (Ala54Pro) adjacent to one of the catalytic residues bestowed hydrolase activity on the modern luciferase by enabling cleavage of this intermediate. Thus, a single substitution next to the catalytic pentad may enable the emergence of promiscuous activity at the enzyme class level, and ancestral reconstruction has a clear potential for obtaining multifunctional catalysts.
ER -
CHALOUPKOVÁ, Radka, Veronika LIŠKOVÁ, Martin TOUL, Klára MARKOVÁ, Eva ŠEBESTOVÁ, Lenka HERNYCHOVÁ, Martin MAREK, José Gaspar RANGEL PAMPLONA PIZARRO PINTO, Daniel PLUSKAL, Jitka WATERMAN, Zbyněk PROKOP and Jiří DAMBORSKÝ. Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts. \textit{ACS Catalysis}. Washington, D.C.: AMER CHEMICAL SOC, 2019, vol.~9, No~6, p.~4810-4823. ISSN~2155-5435. Available from: https://dx.doi.org/10.1021/acscatal.9b01031.
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