Light-Emitting Dehalogenases: Reconstruction of Multifunctional
Biocatalysts

J 2019

Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts

CHALOUPKOVÁ, Radka; Veronika LIŠKOVÁ; Martin TOUL; Klára MARKOVÁ; Eva ŠEBESTOVÁ et. al.

Základní údaje

Originální název

Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts

Autoři

Vydání

ACS Catalysis, Washington, D.C. AMER CHEMICAL SOC, 2019, 2155-5435

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10403 Physical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

Full Text

Impakt faktor

Impact factor: 12.350

Kód RIV

RIV/00216224:14310/19:00107511

Organizační jednotka

Přírodovědecká fakulta

DOI

https://doi.org/10.1021/acscatal.9b01031

UT WoS

000471212600009

EID Scopus

2-s2.0-85065584281

Klíčová slova anglicky

catalytic promiscuity; ancestral reconstruction; haloalkane dehalogenase; monooxygenase; luciferase; emergence of biological function

Štítky

CF PROT, H21, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 15. 2. 2023 22:42, Mgr. Michaela Hylsová, Ph.D.

Anotace

V originále

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.

Návaznosti

EF16_013/0001761, projekt VaV

Název: RECETOX RI

GA16-24223S, projekt VaV

Název: Strukturní podstata vzniku nových enzymových aktivit

Investor: Grantová agentura ČR, Strukturní podstata vzniku nových enzymových aktivit

LM2015043, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

LM2015051, projekt VaV

Název: Centrum pro výzkum toxických látek v prostředí (Akronym: RECETOX RI)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Výzkumná infrastruktura RECETOX

LO1214, projekt VaV

Název: Centrum pro výzkum toxických látek v prostředí (Akronym: RECETOX)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Centrum pro výzkum toxických látek v prostředí

Citovat

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 a Jiří DAMBORSKÝ. Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts. ACS Catalysis. Washington, D.C.: AMER CHEMICAL SOC, 2019, roč. 9, č. 6, s. 4810-4823. ISSN 2155-5435. Dostupné z: https://doi.org/10.1021/acscatal.9b01031.

@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 = {https://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 a Jiří DAMBORSKÝ. Light-Emitting Dehalogenases: Reconstruction of Multifunctional Biocatalysts. \textit{ACS Catalysis}. Washington, D.C.: AMER CHEMICAL SOC, 2019, roč.~9, č.~6, s.~4810-4823. ISSN~2155-5435. Dostupné z: https://doi.org/10.1021/acscatal.9b01031.

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