Bioluminescence emission spectra

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.

Eley, M., et al. (1970). Bacterial bioluminescence. Comparisons of bioluminescence emission spectra, the fluorescence of luciferase reaction mixtures, and the fluorescence of flavin cations. Biochemistry 9 2902-2908. 

Swift, E., Biggley, W. H., and Napora, T. A. (1977). The bioluminescence emission spectra of Pyrosoma atlanticum, P. spinosum (tunicata), Euphausia tenera (Crustacea) and Gonostoma sp. (Pisces)./. Mar. Biol. Assoc. U.K. 57 817-823. 

Widder, E. A., Latz, M. I., and Herring, P. J. (1986). Temporal shifts in bioluminescence emission spectra from the deep-sea fish, Searsia koefoedi. Photochem. Photobiol. 44 97-101. 

EFFECTS OF ORGANIC SOLVENTS ON BIOLUMINESCENCE EMISSION SPECTRA OF BACTERUL LUCIFERASE FROM PHOTOBACTERIUM LEIOGNATHI... 

Figure 1. Bioluminescent emission spectra of bacterial luciferase from P. leiognathi in 1.48 % v/v acetone (1), 0.02 M pH 7.0 phosphate buffer (2) and 2.38 % v/v glycerol (3). Wavelengths of spectral maxima are indicated.

Cline T, Hastings J. Mutated luciferases with altered bioluminescence emission spectra. J Biol Chem 1974 249 4668-9. 

SITE-DIRECTED MUTAGENESIS OF LAMPYRIS TURKESTANICUS LUCIFERASE THE EFFECT OF CONSERVED RESIDUE(S) IN BIOLUMINESCENCE EMISSION SPECTRA AMONG FIREFLY... 

Fig. 1. (A) Partial multiple sequence alignment (for more data refer to the text). (B)The bioluminescence emission spectra produced by the wild-type and mutant luciferases-catalyzed oxidation of luciferin at pH 7.8.

Tafreshi N Kh, Sadeghizadeh M, Emamzadeh R, Ranjbar B, Naderi-Manesh H, Hosseinkhani S. Site-directed mutagenesis of firefly luciferase Implication of conserved residue(s) in bioluminescence emission spectra among firefly luciferases. Biochem J 2008 in press. 

General methods. Luciferase activity, kinetic constants and bioluminescence emission spectra were determined as described previously.2... 

Eley, M., j. Lee, J. M. Lhoste, C. Y. Lee, M. J. Cormier, and P. Hemmerich Bacterial Bioluminescence. Comparisons of Bioluminescence Emission Spectra, the Fluorescence of Luciferase Reaction Mixtures, and the Fluorescence of Flavin Cations. Biochemistry 9, 2902 (1970). 

Eckstein, J. W., et al. (1990). A time-dependent bacterial bioluminescence emission spectrum in an in vitro single turnover system energy transfer alone cannot account for the yellow emission of Vibrio fischeri Y-l. Proc. Natl. Acad. Sci. USA 87 1466-1470. 

The structure of intermediate II is of special interest because its fluorescence emission spectrum after irradiation exactly matches the bioluminescence emission spectrum in the presence of long-chain aldehyde (Fig. 46). In spite of this spectral similarity, it seems almost certain that intermediate II (either before or after irradiation) is not identical with the emitting species, because in order to populate the excited state aldehyde is necessary. Nevertheless, the molecular structure of intermediate II may be of direct relevance to the spectral properties of the emitting species. 

Fig. 3.1.4 Bioluminescence spectrum of Cypridina luciferin catalyzed by Cypridina luciferase (A), the fluorescence excitation spectrum of oxyluciferin in the presence of luciferase (B), the fluorescence emission spectrum of the same solution as B (C), and the absorption spectrum of oxyluciferin (D). The fluorescence of oxyluciferin alone and luciferase alone are negligibly weak. Measurement conditions A, luciferin (lpg/ml) plus a trace amount of luciferase in 20 mM sodium phosphate buffer, pH 7.2, containing 0.2 M NaCl B and C, oxyluciferin (20 pM) plus luciferase (0.2mg/ml) in 20 mM sodium phosphate buffer, pH 7.2, containing 0.2 M NaCl D, oxyluciferin (41 pM) in 20 mM Tris-HCl buffer, pH 7.6, containing 0.2 M NaCl. All are at 20°C.

Fig. 4.1.3 Absorption spectra of aequorin (A), spent solution of aequorin after Ca2+-triggered luminescence (B), and the chromophore of aequorin (C). Fluorescence emission spectrum of the spent solution of aequorin after Ca2+-triggered bioluminescence, excited at 340 nm (D). Luminescence spectrum of aequorin triggered with Ca2+ (E). Curve C is a differential spectrum between aequorin and the protein residue (Shimomura et al., 1974b) protein concentration 0.5 mg/ml for A and B, 1.0 mg/ml for C. From Shimomura and Johnson, 1976.

Fig. 4.2.2 Left panel-. Uncorrected Ca2+-triggered bioluminescence spectrum of W92F obelin derived from O. longissima. Right panel Corrected bioluminescence spectrum of the same obelin (dotted line), and the fluorescence emission spectrum of the spent solution after luminescence (solid line). From Deng et al., 2001, with permission of the Federation of the European Biochemical Societies.

Latia luciferase is colorless and normally nonfluorescent. However, the luciferase fluoresces visibly in alkaline solutions. The fluorescence is most prominent in a KCN solution, showing an emission spectrum that is very close to the bioluminescence spectrum and also to the fluorescence emission of a flavin (FAD) except for the 370 nm... 

Fig. 7.1.5 Fluorescence spectra of purified Chaetopterus photoprotein (CPA) in 10 mM ammonium acetate, pH 6.7 (solid lines), and the bioluminescence spectrum of the luminous slime of Chaetopterus in 10 mM Tris-HCl, pH 7.2 (dashed line). Note that the luminescence spectrum of Chaetopterus photoprotein in 2 ml of 10 mM Tris-HCl, pH 7.2, containing 0.5 M NaCl, 5 pi of old dioxane and 2 pi of 10 mM FeSC>4 (Amax 453-455 nm) matched exactly with the fluorescence emission spectrum of the photoprotein. No significant change was observed in the fluorescence spectrum after the luminescence reaction.

The bioluminescence spectrum of P. stipticus and the fluorescence and chemiluminescence spectra of PM are shown in Fig. 9.7. The fluorescence emission maximum of PM-2 (525 nm) is very close to the bioluminescence emission maximum (530 nm), but the chemiluminescence emission maximum in the presence of a cationic surfactant CTAB (480 nm) differs significantly. However, upon replacing the CTAB with the zwitter-ionic surfactant SB3-12 (3-dodecyldimethylammonio-propanesulfonate), the chemiluminescence spectrum splits into two peaks, 493 nm and 530 nm, of which the latter peak coincides with the emission maximum of the bioluminescence. When PM-1 is heated at 90°C for 3 hr in water containing 10% methanol, about 50% of PM-1 is converted to a new compound that can be isolated by HPLC the chemiluminescence spectrum of this compound in the presence of SB3-12 (curve 5, Fig. 9.7) is practically identical with the bioluminescence spectrum. 

Luminescence of Pyrosoma. All species of the genus Pyrosoma (about 10 species) are bioluminescent. Pyrosoma is one of the few organisms reported to luminesce in response to light (Bowlby et al., 1990). The luminescence emission spectrum of Pyrosoma atlantica is bimodal according to Kampa and Boden (1957), with the primary peak near 482 nm, and the secondary near 525 nm. Swift et al. (1977) reported the emission maxima of two Pyrosoma species at 485 and 493 nm, respectively, and Bowlby et al. (1990) found an emission peak at 475 nm with P. atlantica. A corrected bioluminescence spectrum of P. atlantica (A.max 485 nm) reported by Herring (1983) is shown in Fig. 10.5.2. 

The overall reaction scheme of the luminol chemiluminescence in an aqueous medium is shown in Figure 1. The luminol oxidation leads to the formation of an aminophthalate ion in an excited state, which then emits light on return to the ground state. The quantum yield of the reaction is low ( 0.01) compared with bioluminescence reactions and the emission spectrum shows a maximum1 at 425 nm. 

The dianion 117 bonded to enzyme (firefly luciferase) appears to be the emitter in blue-green firefly bioluminescence as the emission spectrum exactly matches the fluorescence of 117, and in an analogous way in the case of red bioluminescence the emitter is 115. 

Fig. 46. Emission spectrum of bioluminescence (O), measured directly from the cuvette during warming of intermediate II in the presence of aldehyde, plotted togather with the fluorescence emission spectrum (A) of the phototransformed intermediate II. Ordinate intensity of bioluminescence and fluorescence normalized at the peak. From Balny and Hastings (1975). Reprinted with permission of Biochemistry. Copyright by the American Chemical Society.

The emission spectrum of the luminescence reaction of coelenterazine disulfate under the conditions of standard assay (A.max 470 nm) was identical with the luminescence spectrum of homogenate3 and also with the bioluminescence spectrum of the arm light organs. 

The bioluminescence technique is applied for quantitative measurement of tumor burden, treatment response, immune cell trafficking, and detection of gene transfer [40]. The luminescence property depends on adenosine triphosphate and oxygen-dependent enzymatic conversion of exogenous luciferin to oxyluciferin by luciferase within living cells. An emission spectrum is produced with a peak of approximately 560 nm, which can be detected by a highly sensitive charge coupled device camera at 10-15 min after injection of luciferin and lasts for 60 min in mice [41]. 

The emitting molecule, decarboxyketoludferin, has been isolated and synthesized. When it is excited photochemically by photon absorption in basic solution (pH > 7.5-8.0), it fluoresces, giving a fluorescence emission specfrum that is identical to the emission spectrum produced by the interaction of firefly luciferin and firefly luciferase. The emitting form of decarboxykefoluciferin has thus been identified as the enol dianion. In neutral or acidic solution, the emission spectrum of decarboxyketo-luciferin does not match the emission spectrum of the bioluminescent system. 

Bioluminescence of firefly luciferin can produce a wide range of colors when catalyzed by different luciferases obtained from various species of fireflies, with their emission maxima ranging from 535 nm (yellow-green) to 638 nm (red). Apparently, each spectrum is emitted from a single emitting species they are not the composites of the yellow-green peak and the red peak (Seliger and McElroy, 1964). 

The bioluminescence reaction of Oplophorus is a typical luciferin-luciferase reaction that requires only three components luciferin (coelenterazine), luciferase and molecular oxygen. The luminescence spectrum shows a peak at about 454nm (Fig. 3.3.1). The luminescence is significantly affected by pH, salt concentration, and temperature. A certain level of ionic strength (salt) is necessary for the activity of the luciferase. In the case of NaCl, at least 0.05-0.1 M of the salt is needed for a moderate rate of light emission, and about 0.5 M for the maximum light intensity. 

Harvey (1952) demonstrated the luciferin-luciferase reaction with O. phosphorea collected at Nanaimo, British Columbia, Canada, and with O. enopla from Bermuda. McElroy (1960) partially purified the luciferin, and found that the luminescence spectrum of the luciferin-luciferase reaction of O. enopla is identical to the fluorescence spectrum of the luciferin (A.max 510 nm), and also that the luciferin is auto-oxidized by molecular oxygen without light emission. Further investigation on the bioluminescence of Odontosyllis has been made by Shimomura etal. (1963d, 1964) and Trainor (1979). Although the phenomenon is well known, the chemical structure of the luciferin and the mechanism of the luminescence reaction have not been elucidated. 

Fig. 10.1.3 Fluorescence excitation and emission spectra (solid lines) and H2O2-triggered luminescence spectrum (dashed line) of Ophiopsila photoprotein (Shimomura, 1986b, revised). The dotted line indicates the in vivo bioluminescence spectrum of Ophiopsila californica plotted from the data reported by Brehm and Morin (1977).

P. flava emits a green light from whole body by adding diluted H2O2 solution and intermittently continued emitting light around a minute. The bioluminescence spectrum was recorded by using a live specimen and showed emission centered at... 

In thepresence of 4.76 % v/v acetone the luciferase produced emission spectra with increasing maximal emission wavelefagths from 506 to 516 nm (Fig. 2). The methanol-induced bioluminescence change was bimodal with two emission maxima at 502 and 525 nm. In the presence of these methanol concentrations spectrum was a narrow in their spectrum between 450 and 570 nm. 

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