A catalytic mechanism for Renilla -type bioluminescence
A catalytic mechanism for Renilla -type bioluminescence
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 bioluminescence 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 unprecedented snapshots of coelenterazine-to-coelenteramide oxidation. Bound coelenterazine adopts a Y-shaped conformation, enabling the deprotonated imidazopyrazinone component to attack O2 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.
Baatallah Racha、Gagnot Glwadys、Santana Vinicius T.、Janin Yves L.、Schenkmayerova Andrea、Pinto Gaspar P.、Marek Martin、Prokop Zbynek、Neugebauer Petr、Damborsky Jiri、Pluskal Daniel、Stuchla Marketa、Chaiyen Pimchai、Toul Martin、Bednar David
Institut Pasteur, Universit¨| de Paris, CNRS UMR 3523, Unit¨| de Chimie et BiocatalyseInstitut Pasteur, Universit¨| de Paris, CNRS UMR 3523, Unit¨| de Chimie et BiocatalyseCentral European Institute of Technology, CEITEC BUTInstitut Pasteur, Universit¨| de Paris, CNRS UMR 3523, Unit¨| de Chimie et BiocatalyseLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital BrnoLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital BrnoLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital BrnoLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital BrnoCentral European Institute of Technology, CEITEC BUTLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital Brno||Enantis, INBITLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk UniversityLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk UniversitySchool of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC)Loschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital BrnoLoschmidt Laboratories, Department of Experimental Biology and RECETOX, Faculty of Science, Masaryk University||International Clinical Research Center, St. Anne?ˉs University Hospital Brno
生物化学生物物理学生物科学研究方法、生物科学研究技术
bioluminescenceRenilla reniformisluciferasecoelenterazineα/β-hydrolase foldazacoelenterazine
Baatallah Racha,Gagnot Glwadys,Santana Vinicius T.,Janin Yves L.,Schenkmayerova Andrea,Pinto Gaspar P.,Marek Martin,Prokop Zbynek,Neugebauer Petr,Damborsky Jiri,Pluskal Daniel,Stuchla Marketa,Chaiyen Pimchai,Toul Martin,Bednar David.A catalytic mechanism for Renilla -type bioluminescence[EB/OL].(2025-03-28)[2025-07-20].https://www.biorxiv.org/content/10.1101/2022.02.09.479090.点此复制
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