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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