J 2023

Catalytic mechanism for Renilla-type luciferases

SMITH, Andrea, Martin TOUL, Daniel PLUSKAL, Racha BAATALLAH, Glwadys GAGNOT et. al.

Basic information

Original name

Catalytic mechanism for Renilla-type luciferases

Authors

SMITH, Andrea (703 Slovakia, belonging to the institution), Martin TOUL (203 Czech Republic, belonging to the institution), Daniel PLUSKAL (203 Czech Republic, belonging to the institution), Racha BAATALLAH, Glwadys GAGNOT, José Gaspar RANGEL PAMPLONA PIZARRO PINTO (620 Portugal, belonging to the institution), Vinicius T. T. SANTANA, Markéta STUCHLÁ (203 Czech Republic, belonging to the institution), Petr NEUGEBAUER, Pimchai CHAIYEN, Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution), David BEDNÁŘ (203 Czech Republic, belonging to the institution), Yves L. L. JANIN, Zbyněk PROKOP (203 Czech Republic, belonging to the institution) and Martin MAREK (203 Czech Republic, belonging to the institution)

Edition

Nature Catalysis, Nature Portfolio, 2023, 2520-1158

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10403 Physical chemistry

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 37.800 in 2022

RIV identification code

RIV/00216224:14310/23:00130829

Organization unit

Faculty of Science

UT WoS

000906600100001

Keywords in English

CYPRIDINA LUCIFERIN; PROTEIN CRYSTALLIZATION; BIOLUMINESCENCE; RENIFORMIS; LIGHT; CHEMILUMINESCENCE; COELENTERAZINE; STABILITY; LUMINESCENCE; SPECIFICITY

Tags

Tags

International impact, Reviewed
Změněno: 8/4/2024 07:49, Mgr. Marie Šípková, DiS.

Abstract

V originále

The widely used coelenterazine-powered Renilla luciferase was discovered over 40 years ago, but the oxidative mechanism by which it generates blue photons remains unclear. Here we decipher Renilla-type catalysis through crystallographic, spectroscopic and computational experiments. Structures of ancestral and extant luciferases complexed with the substrate-like analogue azacoelenterazine or a reaction product were obtained, providing molecular snapshots of coelenterazine-to-coelenteramide oxidation. Bound coelenterazine adopts a Y-shaped conformation, enabling the deprotonated imidazopyrazinone component to attack O-2 via a radical charge-transfer mechanism. A high emission intensity is secured by an aspartate from a conserved proton-relay system, which protonates the excited coelenteramide product. Another aspartate on the rim of the catalytic pocket fine-tunes the electronic state of coelenteramide and promotes the formation of the blue light-emitting phenolate anion. The results obtained also reveal structural features distinguishing flash-type from glow-type bioluminescence, providing insights that will guide the engineering of next-generation luciferase-luciferin pairs for ultrasensitive optical bioassays.

Links

EF17_043/0009632, research and development project
Name: CETOCOEN Excellence
GA22-09853S, research and development project
Name: Analýza vzniku nových proteinových funkcí v rámci rodiny halogenalkan dehalogenas
Investor: Czech Science Foundation
LM2018121, research and development project
Name: Výzkumná infrastruktura RECETOX (Acronym: RECETOX RI)
Investor: Ministry of Education, Youth and Sports of the CR, RECETOX RI
LM2018127, research and development project
Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
LM2018140, research and development project
Name: e-Infrastruktura CZ (Acronym: e-INFRA CZ)
Investor: Ministry of Education, Youth and Sports of the CR
MUNI/H/1561/2018, interní kód MU
Name: Decoding the molecular principles of enzyme evolution
Investor: Masaryk University, Individual High risk/high gain projects
792772, interní kód MU
Name: Structural and biochemical studies of an ancestral enzyme with dual dehalogenase and luciferase activity (Acronym: Ancestral)
Investor: European Union, MSCA Marie Skłodowska-Curie Actions (Excellent Science)
814418, interní kód MU
Name: Synthetic biology-guided engineering of Pseudomonas putida for biofluorination (Acronym: SinFonia)
Investor: European Union, Leadership in enabling and industrial technologies (LEIT) (Industrial Leadership)
857560, interní kód MU
(CEP code: EF17_043/0009632)
Name: CETOCOEN Excellence (Acronym: CETOCOEN Excellence)
Investor: European Union, Spreading excellence and widening participation
90242, large research infrastructures
Name: CIISB III