Publication detail

Catalytic mechanism for Renilla-type luciferases

Schenkmayerova, A. Toul, M. Pluskal, D. Baatallah, R. Gagnot, G. Pinto, GP. Santana, VT. Stuchla, M. Neugebauer, P. Chaiyen, P. Damborsky, J. Bednar, D. Janin, YL. Prokop, Z. Marek, M.

Original Title

Catalytic mechanism for Renilla-type luciferases

Type

journal article in Web of Science

Language

English

Original Abstract

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.

Keywords

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

Authors

Schenkmayerova, A.; Toul, M.; Pluskal, D.; Baatallah, R.; Gagnot, G.; Pinto, GP.; Santana, VT.; Stuchla, M.; Neugebauer, P.; Chaiyen, P.; Damborsky, J.; Bednar, D.; Janin, YL.; Prokop, Z.; Marek, M.

Released

1. 1. 2023

Publisher

NATURE PORTFOLIO

Location

BERLIN

ISBN

2520-1158

Periodical

Nature Catalysis

Year of study

6

Number

1

State

United Kingdom of Great Britain and Northern Ireland

Pages from

23

Pages to

38

Pages count

16

URL

https://www.nature.com/articles/s41929-022-00895-z

BibTex

@article{BUT183685,
  author="Schenkmayerova, A. and Toul, M. and Pluskal, D. and Baatallah, R. and Gagnot, G. and Pinto, GP. and Santana, VT. and Stuchla, M. and Neugebauer, P. and Chaiyen, P. and Damborsky, J. and Bednar, D. and Janin, YL. and Prokop, Z. and Marek, M.",
  title="Catalytic mechanism for Renilla-type luciferases",
  journal="Nature Catalysis",
  year="2023",
  volume="6",
  number="1",
  pages="23--38",
  doi="10.1038/s41929-022-00895-z",
  issn="2520-1158",
  url="https://www.nature.com/articles/s41929-022-00895-z"
}