1.13.12.5 12-benzyl-8-hydroxy-2-(4-hydroxybenzyl)-5,11-dihydrobenzo[f]imidazo[1,2-a]quinoxalin-3(6H)-one + O2 - Renilla reniformis oxidized 12-benzyl-8-hydroxy-2-(4-hydroxybenzyl)-5,11-dihydrobenzo[f]imidazo[1,2-a]quinoxalin-3(6H)-one + CO2 + hnu - ? 372482 1.13.12.5 2,8-dibenzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis oxidized 2,8-dibenzyl-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + CO2 + hn - ? 372484 1.13.12.5 2-benzyl-3-[(2-nitrophenyl)methoxy]-6-phenyl-8-(phenylsulfanyl)imidazo[1,2-a]pyrazine + O2 the substrate presents robust bioluminescent signals ex vivo and in living animals after UV irradiation at 365 nm Renilla sp. ? + CO2 - ? 461727 1.13.12.5 2-benzyl-3-[(4,5-dimethoxy-2-nitrophenyl)methoxy]-6-phenyl-8-(phenylsulfanyl)imidazo[1,2-a]pyrazine + O2 the substrate presents robust bioluminescent signals ex vivo and in living animals after UV irradiation at 365 nm Renilla sp. ? + CO2 - ? 461728 1.13.12.5 2-benzyl-3-[1-(2-nitrophenyl)ethoxy]-6-phenyl-8-(phenylsulfanyl)imidazo[1,2-a]pyrazine + O2 the substrate presents robust bioluminescent signals ex vivo and in living animals after UV irradiation at 365 nm Renilla sp. ? + CO2 - ? 461729 1.13.12.5 2-benzyl-8-benzyl-6-(2-fluorophenylethynyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442766 1.13.12.5 2-benzyl-8-benzyl-6-(3-fluorophenylethynyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442767 1.13.12.5 2-benzyl-8-benzyl-6-(3-hydroxyphenylethynyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442768 1.13.12.5 2-benzyl-8-benzyl-6-(3-methylphenylethynyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442769 1.13.12.5 2-benzyl-8-benzyl-6-(4-fluorophenylethynyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442770 1.13.12.5 2-benzyl-8-benzyl-6-(phenylethynyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442771 1.13.12.5 2-benzyl-8-benzyl-6-[(1-fluoroethyl)-1,2,3-triazol-4]imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442772 1.13.12.5 2-benzyl-8-benzyl-6-[(1-hydroxyethyl)-1,2,3-triazol-4]imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442773 1.13.12.5 2-benzyl-8-benzyl-6-[(1-hydroxypropyl)-1,2,3-triazol-4]imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis ? + CO2 + hv - ? 442774 1.13.12.5 2-hydroperoxycoelenterazine + O2 - Aequorea victoria ? - ? 387140 1.13.12.5 3iso-coelenterazine + O2 relative activity to native coelenterazine: 11.6% Renilla reniformis ? - ? 428009 1.13.12.5 3iso-coelenterazine + O2 relative activity to native coelenterazine: 14.3% Gaussia ? - ? 428009 1.13.12.5 3iso-coelenterazine + O2 relative activity to native coelenterazine: 34.9% Renilla reniformis ? - ? 428009 1.13.12.5 3iso-coelenterazine + O2 relative activity to native coelenterazine: 78.2% Oplophorus gracilirostris ? - ? 428009 1.13.12.5 3me-coelenterazine + O2 relative activity to native coelenterazine: 11.8% Renilla reniformis ? - ? 428011 1.13.12.5 3me-coelenterazine + O2 relative activity to native coelenterazine: 2.6% Gaussia ? - ? 428011 1.13.12.5 3me-coelenterazine + O2 relative activity to native coelenterazine: 45.5% Renilla reniformis ? - ? 428011 1.13.12.5 3me-coelenterazine + O2 relative activity to native coelenterazine: 80% Oplophorus gracilirostris ? - ? 428011 1.13.12.5 3meo-coelenterazine + O2 relative activity to native coelenterazine: 189% Oplophorus gracilirostris ? - ? 428013 1.13.12.5 3meo-coelenterazine + O2 relative activity to native coelenterazine: 3.8% Gaussia ? - ? 428013 1.13.12.5 3meo-coelenterazine + O2 relative activity to native coelenterazine: 5.5% Renilla reniformis ? - ? 428013 1.13.12.5 3meo-coelenterazine + O2 relative activity to native coelenterazine: 82.3% Renilla reniformis ? - ? 428013 1.13.12.5 8-benzyl-2-(4-fluorobenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis oxidized 8-benzyl-2-(4-fluorobenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + CO2 + hnu - ? 372483 1.13.12.5 8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + O2 - Renilla reniformis oxidized 8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one + CO2 + hn - ? 372485 1.13.12.5 alphameh-coelenterazine + O2 relative activity to native coelenterazine: 0.02% Renilla reniformis ? - ? 428159 1.13.12.5 alphameh-coelenterazine + O2 relative activity to native coelenterazine: 0.02% Gaussia ? - ? 428159 1.13.12.5 alphameh-coelenterazine + O2 relative activity to native coelenterazine: 15.7% Oplophorus gracilirostris ? - ? 428159 1.13.12.5 alphameh-coelenterazine + O2 relative activity to native coelenterazine: 24.7% Renilla reniformis ? - ? 428159 1.13.12.5 b-coelenterazine + O2 - Renilla sp. ? - ? 404486 1.13.12.5 beetle D-luciferin 450 microM Gaussia princeps ? - ? 404456 1.13.12.5 benzylluciferin + O2 - Renilla reniformis oxidized benzylluciferin + CO2 + hv - ? 288827 1.13.12.5 benzylluciferin methyl ether + O2 - Renilla reniformis oxidized benzylluciferin methyl ether + CO2 + hv - ? 288828 1.13.12.5 bis-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 404541 1.13.12.5 cf3-coelenterazine + O2 relative activity to native coelenterazine: 0.8% Gaussia ? - ? 428481 1.13.12.5 cf3-coelenterazine + O2 relative activity to native coelenterazine: 16.8% Renilla reniformis ? - ? 428481 1.13.12.5 cf3-coelenterazine + O2 relative activity to native coelenterazine: 49.5% Oplophorus gracilirostris ? - ? 428481 1.13.12.5 cf3-coelenterazine + O2 relative activity to native coelenterazine: 5% Renilla reniformis ? - ? 428481 1.13.12.5 coelenterate-type luciferin + O2 - Renilla reniformis oxidized coelenterate-type luciferin + CO2 + hv - ? 288826 1.13.12.5 coelenterazine + ? 20 microM Gaussia princeps ? - ? 404687 1.13.12.5 coelenterazine + O2 - Renilla reniformis oxidized coelenterazine + CO2 + hn - ? 372744 1.13.12.5 coelenterazine + O2 - Renilla sp. ? - ? 388389 1.13.12.5 coelenterazine + O2 - Aequorea forskalea ? - ? 388389 1.13.12.5 coelenterazine + O2 - Gaussia princeps ? - ? 388389 1.13.12.5 coelenterazine + O2 relative activity: 100% Renilla reniformis ? - ? 388389 1.13.12.5 coelenterazine + O2 relative activity: 100% Oplophorus gracilirostris ? - ? 388389 1.13.12.5 coelenterazine + O2 relative activity: 100% Gaussia ? - ? 388389 1.13.12.5 coelenterazine + O2 0.1 mM Gaussia princeps ? - ? 388389 1.13.12.5 coelenterazine + O2 20 microM, assay at pH 8.0 Gaussia princeps ? - ? 388389 1.13.12.5 coelenterazine + O2 assay at pH 7.2 Renilla reniformis ? - ? 388389 1.13.12.5 coelenterazine + O2 - Renilla reniformis coelenteramide + CO2 + hv - ? 388390 1.13.12.5 coelenterazine + O2 - Clytia gregaria coelenteramide + CO2 + hv - ? 388390 1.13.12.5 coelenterazine + O2 blue fluorescent protein from the calcium-binding photoprotein aequorin (BFP-aq) is a complex of Ca2+-bound apoaequorin and coelenteramide, and shows luminescence activity like a luciferase. The oxidation of coelenterazine by BFP-aq in the luciferase reaction and the regeneration process to aequorin might involve the same catalytic site of BFP-aq Aequorea forskalea coelenteramide + CO2 + hv - ? 388390 1.13.12.5 coelenterazine + O2 clytin emits light on reacting with Ca2+. The Ca2+ sensitivity of recombinant clytin is lower than that of aequorin Clytia gregaria coelenteramide + CO2 + hv - ? 388390 1.13.12.5 coelenterazine + O2 coelenterazine is bound by coelenterazine-binding protein. In combinantion with renilla luciferase, addition of only one Ca2+-ion is sufficient to release coelenterazine as a substrate. The combination of the two proteins generates bioluminescence with higher reaction efficiency than using free coelenterazine alone Renilla muelleri coelenteramide + CO2 + hv - ? 388390 1.13.12.5 coelenterazine + O2 - Renilla sp. coelenteramide + CO2 + hnu - ? 404688 1.13.12.5 coelenterazine + O2 - Gaussia princeps coelenteramide + CO2 + hnu - ? 404688 1.13.12.5 coelenterazine + O2 0.5 microg, assay at pH 7.6, high substrate specificity for coelenterazine Gaussia princeps coelenteramide + CO2 + hnu - ? 404688 1.13.12.5 coelenterazine + O2 assay at pH 7.4, 25-30°C, 10 min Gaussia princeps coelenteramide + CO2 + hnu - ? 404688 1.13.12.5 coelenterazine + O2 the enzyme catalyzes the oxidative decarboxylation of coelenterazine in the presence of O2, resulting in the formation of coelenteramide in its excited state and CO2 as the reaction product Renilla muelleri coelenteramide + CO2 + hnu green bioluminescence ? 404688 1.13.12.5 coelenterazine + O2 - Gaussia princeps ? + CO2 + hv - ? 462369 1.13.12.5 coelenterazine h + O2 - Renilla reniformis coelenteramide h + CO2 + hv - ? 443389 1.13.12.5 coelenterazine h + O2 - Renilla reniformis excited coelenteramide h monoanion + CO2 - ? 461575 1.13.12.5 coelenterazine h + O2 an induced-fit mechanism is proposed where ligand-binding induces conformational changes of the active site. Insights regarding the controversial properties and the mechanism of the reaction catalysis of Renilla luciferase and its red-shifted light emittingvariant (Super RLuc 8) Renilla reniformis excited coelenteramide h monoanion + CO2 - ? 461575 1.13.12.5 coelenterazine-H + O2 - Renilla reniformis ? - ? 404689 1.13.12.5 coelenterazine-H + O2 5 microM, assay at pH 7.5 Renilla reniformis ? - ? 404689 1.13.12.5 coelenterazine-h + O2 substrate binding structure Renilla reniformis coelenteramide h + CO2 + hv - ? 443390 1.13.12.5 coelenterazine-v + O2 increase of substrate coelenterazine stability by ligating it to Ca2+-triggered coelenterazine-binding protein, CBP, from Renilla muelleri, which apparently functions in the organism for stabilizing and protecting coelenterazine from oxidation. The apo-CBP binds coelenterazine-v very rapidly from Ca2+ free solution, similar to that as the native coelenterazine. At low concentrations, coelenterazine-v bound within CBP generates a brighter bioluminescence signal than would free coelenterazine, thereby increasing the assay sensitivity, overview Renilla muelleri coelenteramide-v + CO2 + hnu orange bioluminescence ? 414448 1.13.12.5 coelentrazine + O2 - Gaussia princeps ? - ? 404690 1.13.12.5 cp-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 404709 1.13.12.5 D-luciferin + O2 - Renilla sp. ? - ? 404867 1.13.12.5 D-luciferin + O2 - Gaussia princeps ? - ? 404867 1.13.12.5 D-luciferin + O2 + ATP - Renilla reniformis oxidized D-luciferin + CO2 + H2O + AMP + diphosphate + hv - ? 388455 1.13.12.5 e-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 405054 1.13.12.5 et-coelenterazine + O2 relative activity to native coelenterazine: 0.9% Gaussia ? - ? 428607 1.13.12.5 et-coelenterazine + O2 relative activity to native coelenterazine: 1.2% Renilla reniformis ? - ? 428607 1.13.12.5 et-coelenterazine + O2 relative activity to native coelenterazine: 21.1% Oplophorus gracilirostris ? - ? 428607 1.13.12.5 et-coelenterazine + O2 relative activity to native coelenterazine: 78.3% Renilla reniformis ? - ? 428607 1.13.12.5 f-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 405108 1.13.12.5 fcp-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 405121 1.13.12.5 h-coelenterazine + O2 assay at pH 7.6m, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 405399 1.13.12.5 h-coelenterazine + O2 relative activity to native coelenterazine: 12.4% Renilla reniformis ? - ? 428670 1.13.12.5 h-coelenterazine + O2 relative activity to native coelenterazine: 5.8% Gaussia ? - ? 428670 1.13.12.5 h-coelenterazine + O2 relative activity to native coelenterazine: 54.1% Renilla reniformis ? - ? 428670 1.13.12.5 h-coelenterazine + O2 relative activity to native coelenterazine: 68.4% Oplophorus gracilirostris ? - ? 428670 1.13.12.5 hcp-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 405406 1.13.12.5 i-coelenterazine + O2 relative activity to native coelenterazine: 0.3% Renilla reniformis ? - ? 428699 1.13.12.5 i-coelenterazine + O2 relative activity to native coelenterazine: 0.9% Gaussia ? - ? 428699 1.13.12.5 i-coelenterazine + O2 relative activity to native coelenterazine: 32.3% Oplophorus gracilirostris ? - ? 428699 1.13.12.5 i-coelenterazine + O2 relative activity to native coelenterazine: 5.1% Renilla reniformis ? - ? 428699 1.13.12.5 luciferin + O2 assay at pH 7.4, 25-30°C, 10 min Gaussia princeps ? - ? 405739 1.13.12.5 me-coelenterazine + O2 relative activity to native coelenterazine: 4.5% Gaussia ? - ? 428783 1.13.12.5 me-coelenterazine + O2 relative activity to native coelenterazine: 46.6% Oplophorus gracilirostris ? - ? 428783 1.13.12.5 me-coelenterazine + O2 relative activity to native coelenterazine: 5.2% Renilla reniformis ? - ? 428783 1.13.12.5 me-coelenterazine + O2 relative activity to native coelenterazine: 73.6% Renilla reniformis ? - ? 428783 1.13.12.5 MeO-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 405814 1.13.12.5 meo-coelenterazine + O2 relative activity to native coelenterazine: 1.2% Renilla reniformis ? - ? 428785 1.13.12.5 meo-coelenterazine + O2 relative activity to native coelenterazine: 13.6% Gaussia ? - ? 428785 1.13.12.5 meo-coelenterazine + O2 relative activity to native coelenterazine: 18.2% Renilla reniformis ? - ? 428785 1.13.12.5 meo-coelenterazine + O2 relative activity to native coelenterazine: 68.1% Oplophorus gracilirostris ? - ? 428785 1.13.12.5 methyl luciferin + O2 - Renilla reniformis ? - ? 453847 1.13.12.5 additional information 8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo-[1,2-a]pyrazin-3 (7H)-one is similar to, and as active as Renilla luciferin, the native luciferin has the benzyl group replaced by an unidentified group of approximately 200 Da Renilla reniformis ? - ? 89 1.13.12.5 additional information native 8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one and its derivatives are most promising substrates for use with Renilla luciferin reporter enzyme in cell culture and living animals Renilla reniformis ? - ? 89 1.13.12.5 additional information free coelenterazine and its analogues are unstable in neutral aqueous solution, undergoing a slow autooxidation over several hours Renilla muelleri ? - ? 89 1.13.12.5 additional information design and synthesis of bioluminescent coelenterazine derivatives (alkynes and triazoles) with imidazopyrazinone C-6 extended substitution as substrates for Renilla luciferase, evaluation, substrate specificity, overview Renilla reniformis ? - ? 89 1.13.12.5 additional information dynamic light scattering has been used for the detection of protein-protein interaction between the IgG antibody and modified enzyme FcUni-RLuc, to which an Fc-binding peptide is bound and separated by a five-amino acid linker from RLuc. Analysis of FcUni-RLuc and Herceptin interaction Renilla reniformis ? - ? 89 1.13.12.5 additional information establishment and evaluation of an indirect autophagy flux assay based on monitoring the degradation of an autophagosome-associated fusion protein Rluc-LC3 by luminescence detection. The Rluc-LC3 assay is useful for the identification of genes, miRNAs, and small molecules that regulate autophagy flux in mammalian cells. LC3 is a subfamily in the Atg8 family of ubiquitin-like proteins and is the only protein marker described to reliably bind the autophagosomal membrane and the phagophore Method evaluation and optimization, overview Renilla reniformis ? - ? 89 1.13.12.5 additional information substrate of Renilla luciferase, coelenterazine, is a heterocyclic imidazolo-pyrazinone, which is derivatized with (4-hydroxyphenyl)methyl (R2), 4-hydroxyphenyl (R6), and phenyl-methyl (R8) moieties Renilla reniformis ? - ? 89 1.13.12.5 n-coelenterazine + O2 assay at pH 7.6, about 100fold reduced relative luminescence Gaussia princeps hnu + ? - ? 406000 1.13.12.5 Renilla luciferin + O2 - Renilla reniformis oxidized Renilla luciferin + CO2 + hv - ? 288825 1.13.12.5 Renilla luciferin + O2 - Renilla muelleri oxidized Renilla luciferin + CO2 + hv - ? 288825 1.13.12.5 Renilla luciferin + O2 - Renilla koellikeri oxidized Renilla luciferin + CO2 + hv - ? 288825