Coelenterazine 400a, a derivative of natural coelenterazine[1], functions as a substrate for Renilla luciferase (Rluc)[2], with a maximum emission wavelength of 400nm. In the presence of Rluc, Coelenterazine 400a reacts with molecular oxygen, forming high-energy intermediates and emitting light as a result[3][4]. Its minimal interference with fluorescent acceptors makes it an ideal choice for bioluminescence resonance energy transfer (BRET) assays, commonly used to evaluate protein-protein interactions[5][6].
References:
[1]. Sousa J, Magalhães CM, González-Berdullas P, et al. Comparative Investigation of the Chemiluminescent Properties of a Dibrominated Coelenterazine Analog. Int J Mol Sci. 2022 Jul 30;23(15):8490. doi: 10.3390/ijms23158490. PMID: 35955625; PMCID: PMC9369366.
[2]. Markova SV, Vysotski ES. Coelenterazine-dependent luciferases. Biochemistry (Mosc). 2015 Jun;80(6):714-32. doi: 10.1134/S0006297915060073. PMID: 26531017.
[3]. Min CG, Ferreira PJO, Pinto da Silva L. Theoretically obtained insight into the mechanism and dioxetanone species responsible for the singlet chemiexcitation of Coelenterazine. J Photochem Photobiol B. 2017 Sep;174:18-26. doi: 10.1016/j.jphotobiol.2017.07.012. Epub 2017 Jul 20. PMID: 28750319.
[4]. Vacher M, Fdez Galván I, Ding BW, et al. Chemi- and Bioluminescence of Cyclic Peroxides. Chem Rev. 2018 Aug 8;118(15):6927-6974. doi: 10.1021/acs.chemrev.7b00649. Epub 2018 Mar 1. PMID: 29493234.
[5]. Ming-Liang Yuan, Tian-Yu Jiang, Lu-Pei Du, et al. Luminescence of coelenterazine derivatives with C-8 extended electronic conjugation. Chinese Chemical Letters. 2016, 27(04): 550-554 https://doi.org/10.1016/j.cclet.2016.02.011
[6]. Li J, Wang X, Dong G, et al. Novel furimazine derivatives for nanoluciferase bioluminescence with various C-6 and C-8 substituents. Org Biomol Chem. 2021 Sep 22;19(36):7930-7936. doi: 10.1039/d1ob01098k. PMID: 34549229.
Coelenterazine 400a是一种天然腔肠素的衍生物[1]，作为海肾荧光素酶（Rluc）的底物[2]，其最大发射波长约为400nm。在Rluc的存在下Coelenterazine 400a与分子氧反应，形成高能量的中间产物，并发出光[3][4]。由于其对荧光受体的干扰极小，使其成为生物发光共振能量转移（BRET）实验的理想底物，常用于评估蛋白质-蛋白质相互作用[5][6]。