Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications

Abstract

: The functioning of bioluminescent systems in most of the known marine organisms is based on the oxidation reaction of the same substrate—coelenterazine (CTZ), catalyzed by luciferase. Despite the diversity in structures and the functioning mechanisms, these enzymes can be united into a common group called CTZ-dependent luciferases. Among these, there are two sharply different types of the system organization—Ca2+-regulated photoproteins and luciferases themselves that function in accordance with the classical enzyme–substrate kinetics. Along with deep and comprehensive fundamental research on these systems, approaches and methods of their practical use as highly sensitive reporters in analytics have been developed. The research aiming at the creation of artificial luciferases and synthetic CTZ analogues with new unique properties has led to the development of new experimental analytical methods based on them. The commercial availability of many ready-to-use assay systems based on CTZ-dependent luciferases is also important when choosing them by first-time-users. The development of analytical methods based on these bioluminescent systems is currently booming. The bioluminescent systems under consideration were successfully applied in various biological research areas, which confirms them to be a powerful analytical tool. In this review, we consider the main directions, results, and achievements in research involving these luciferases.

How to Select Firefly Luciferin Analogues for In Vivo Imaging

“…This reaction proceeds by two-steps: DLH2 is first adenylated with ATP, and then it is oxidized by O2, to generate excited oxyluciferin, which produces yellow-green light (λmax = 560 nm) [4][5][6][7]. In contrast, bioluminescence in the luminous shrimp, one of many bioluminescent marine species, is generated by the oxidation of coelenterazine (CTZ) catalyzed by oplophorus luciferase (Oluc) in the presence of O2 to generate coelenteramide, which produces blue light (λmax = 454 nm) [8][9][10]. [1], [2], [3], [4], [5], [6], [ Gaussia luciferase (Gluc) Gaussia princeps O2 [1], [2], [3], [8], [9], [10] Renilla luciferase (Rluc)…”

Molecular Design of d-Luciferin-Based Bioluminescence and 1,2-Dioxetane-Based Chemiluminescence Substrates for Altered Output Wavelength and Detecting Various Molecules

“…In this review, a summary is presented of the reported molecular design of substrates emitting NIR-I bioluminescence and probes with a focus on the firefly/click beetle luciferin-luciferase system. Similar studies have been carried out in a bioluminescent system using coelenterazine as a substrate, but refer to other excellent reviews [10,11]. There is also a bioluminescent system with bacterial luciferinluciferase reaction, but it is not discussed in this review because it does not require the addition of luciferin from outside; therefore, there is no research on the molecular design of bacterial luciferin [12,13].…”

H2O-Bridged Proton-Transfer Channel in Emitter Species Formation in Obelin Bioluminescence

“…Moreover, in contrast to luciferases, the CaRP light emission reaction does not require molecular oxygen and CaRP reacts only once, i.e., the one does not “turn over” as an enzyme does . Their broad analytical potential originating from their ability to emit light on Ca 2+ addition causes broad interest in these photoproteins. − …”