Renilla, bioluminescence

Fig. 4.6.2 A scheme showing the mechanism of the Renilla bioluminescence, and the chemical structures of coelenterazine derivatives involved.

Anderson, J. M., Charbonneau, H., and Cormier, M. J. (1974). Mechanism of calcium induction of Renilla bioluminescence. Involvement of a calcium-triggered luciferin binding protein. Biochemistry 13 1195-1200. 

Renilla, bioluminescence, 1255 Resomfin, hydrogen peroxide determination, 642-3... 

The product coelenteramide is not noticeably fluorescent in aqueous solutions, but is highly fluorescent in organic solvents and also when the compound is in the hydrophobic environment of a protein. When coelenterazine is luminesced in the presence of Oplophorus luciferase, the solution after luminescence (the spent solution) is not fluorescent, presumably due to the dissociation of coelenteramide from the luciferase that provided a hydrophobic environment at the time of light emission. An analogous situation exists in the bioluminescence system of Renilla (Hori et al., 1973). 

Anthozoa. Anthozoans are plant-shaped polyps, either solitary or colonial, completely lacking the medusoid stage. They are found along coastal waters and include the luminescent genera Renilla (the sea pansies), Cavernularia (the sea cactuses), and Ptilosarcus and Pennatula (the sea pens). Bioluminescent anthozoans emit light by a luciferin-luciferase reaction that involves coelenterazine as the... 

Fig. 4.6.3 Bioluminescence emission spectra measured with coelenterazine plus 1 i.M Renilla luciferase in the absence (a) and presence (b) of 1 jlM Renilla GFP. From Lorenz et al., 1991.

Soon after the hypothetical structure was published, coelenterazine was isolated as an actual substance from the liver of the luminous squid Watasenia scintillans, and it was chemically synthesized (Inoue et al., 1975). The availability of synthetic coelenterazine led to the important discovery that the treatment of the luminescence product of aequorin with coelenterazine results in the regeneration of active aequorin (Shimomura and Johnson, 1975c), which consequently confirmed the presence of a coelenterazine moiety in the aequorin molecule. During the same period, it became increasingly evident that coelenterazine is involved as a luciferin in various bioluminescent organisms, such as the sea cactus Cavernularia, the sea pen Ptilosarcus, and the sea pansy Renilla (Shimomura and Johnson, 1975b). 

Coelenterazine emits chemiluminescence when dissolved in dimethyl sulfoxide (DMSO) or dimethylformamide (DMF) containing a trace amount of base. It also emits bioluminescence in aqueous media in the presence of a coelenterazine luciferase, such as Renilla luciferase or Oplophorus luciferase. In both cases, the luminescence reactions require molecular oxygen. The capability of coelenterazine to produce luminescence is attributed to the presence of the imida-zopyrazinone structure in the molecule. 

Based on the available knowledge on the chemiluminescence and bioluminescence reactions of various luciferins (firefly, Cypridina, Oplophorus and Renilla), the luminescence reaction of coelenterazine is considered to proceed as shown in Fig. 5.4 (p. 171). The reaction is initiated by the binding of O2 at the 2-position of the coelenterazine molecule, giving a peroxide. The peroxide then forms a four-membered ring dioxetanone, as in the case of the luminescence... 

The enol-sulfate form (I), which is the precursor of the luciferin in the bioluminescence system of the sea pansy Renilla (Hori et al., 1972), can be readily converted into coelenterazine by acid hydrolysis. The enol-sulfate (I), dehydrocoeienterazine (D) and the coelenterazine bound by the coelenterazine-binding proteins are important storage forms for preserving unstable coelenterazine in the bodies of luminous organisms. The disulfate form of coelenterazine (not shown in Fig. 5.5) is the luciferin in the firefly squid Watasenia (Section 6.3.1). An enol-ether form of coelenterazine bound with glucopyra-nosiduronic acid has been found in the liver of the myctophid fish Diapbus elucens (Inoue et al., 1987). 

Cormier, M. J., and Dure, L. S. (1963). Studies on the bioluminescence of Balanoglossus biminiensis extracts. I. Requirement for hydrogen peroxide and characteristics of the system. J. Biol. Chem. 238 785-789. Cormier, M. J., and Hori, K. (1963). Studies on the bioluminescence of Renilla reniformis. IV. Non-enzymatic activation of Renilla luciferin. Biochim. Biophys. Acta 88 99-104. 

DeLuca, M., et al. (1971). Mechanism of oxidative carbon dioxide production during Renilla reniformis bioluminescence. Proc. Natl. Acad. Sci. USA 68 1658-1660. 

Hart, R. C., Matthews, J. C., Hori, K., and Cormier, M. J. (1979). Renilla reniformis bioluminescence Luciferase-catalyzed production of nonradiating excited states from luciferin analogues and elucidation of the excited state species involved in energy transfer to Renilla green fluorescent protein. Biochemistry 18 2204-2210. 

Hastings, J. W., and Morin, J. C. (1969). Calcium-triggered light emission in Renilla. A unitary biochemical scheme for coelenterate bioluminescence. Biochem. Biophys. Res. Commun. 37 493-498. 

Inouye, S., and Shimomura, O. (1997). The use of Renilla luciferase, Oplophorus luciferase, and apoaequorin as bioluminescent reporter protein in the presence of coelenterazine analogues as substrate. Biochem. Biophys. Res. Commun. 233 349-353. 

Kreiss, P., and Cormier, M. J. (1967). Inhibition of Renilla reniformis bioluminescence by light effects on luciferase and its substrates. Biochim. Biophys. Acta 141 181-183. 

Ward WW, Cormier MJ (1979) An energy transfer protein in coelenterate bioluminescence. Characterization of the Renilla green-fluorescent protein. J Biol Chem 254 781-788... 

Bioluminescence studies research and resources by John E. Wampler (Renilla green fluorescent protein and other organisms)... 

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