Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00209805%3A_____%2F19%3A00078138" target="_blank" >RIV/00209805:_____/19:00078138 - isvavai.cz</a>
Alternative codes found
RIV/00216224:14310/19:00107511 RIV/00159816:_____/19:00071064
Result on the web
<a href="https://pubs.acs.org/doi/full/10.1021/acscatal.9b01031" target="_blank" >https://pubs.acs.org/doi/full/10.1021/acscatal.9b01031</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1021/acscatal.9b01031" target="_blank" >10.1021/acscatal.9b01031</a>
Alternative languages
Result language
angličtina
Original language name
Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts
Original language description
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 Å 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 hydrogenMINUS SIGN 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.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2019
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
ACS catalysis
ISSN
2155-5435
e-ISSN
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Volume of the periodical
2019
Issue of the periodical within the volume
9
Country of publishing house
US - UNITED STATES
Number of pages
14
Pages from-to
"4810−4823"
UT code for WoS article
000471212600009
EID of the result in the Scopus database
2-s2.0-85065584281