Luciferin-luciferase reaction

The luciferin-luciferase reaction of fireflies was first demonstrated by Harvey (1917), although the light observed was weak and short-lasting. Thirty years after Harvey s discovery, McElroy (1947) made a crucial breakthrough in the study of firefly bioluminescence. He found that the light-emitting reaction requires ATP as a cofactor. The addition of ATP to the mixtures of luciferin and luciferase... 

The elaterid Pyrophorus is of special importance in the history of bioluminescence, because it was used by Dubois in his first demonstration of the luciferin-luciferase reaction in 1885. The Jamaican click beetle (Pyrophorus noctilucus) is commonly found in the West Indies. The beetle possesses two kinds of luminous organs. A... 

The luciferin-luciferase reaction of Arachnocampa was first demonstrated by Wood (1993), by mixing a cold-water extract and a cooled hot-water extract. The cold-water extract was prepared with 27 mM Tricine, pH 7.4, containing 7mM MgSC>4, 0.2 mM EDTA, 10% glycerol and 1% Triton X-100, and incubated with 1 mM ATP on ice for 18 hr. The hot-water extract was prepared by heating the cold water extract before the addition of ATP at 98°C for 5 min. The luminescence reaction was performed in the presence of 1 mM ATP. 

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. 

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... 

The scyphozoan Periphylla emits light with a luciferin-luciferase reaction using coelenterazine as the luciferin, differing from Pelagia in the same class and all luminous hydrozoans that luminesce with photoproteins. 

Bivalvia. The bivalve Pholas is historically important because the concept of luciferin-luciferase reaction was established with this clam (Dubois, 1887). It is the only bivalve that is well known and biochemically investigated. The details of the Pholas bioluminescence are given in Section 6.2. 

The New Zealand freshwater limpet Latia neritoides (Fig. 6.1.1) is the only known example of a freshwater luminous organism, with two possible exceptions certain species of luminous bacteria and the larvae of certain species of fireflies. The limpet inhabits shallow clear streams in the North Island of New Zealand, clinging to stones and rocks. Latia has a small oval-shaped shell (6-8 mm long), and secretes a luminous mucus that emits a greenish glow around the body only when disturbed the limpet does not show a spontaneous luminescence. The luminescence of Latia was first reported by Suter (1890) and further details including a positive luciferin-luciferase reaction were described by Bowden (1950). Both the luciferin and the luciferase have... 

Kojima et al. (2000a) that the purified luciferase exhibited a luciferin-luciferase reaction with Latia luciferin without any cofactor. Nevertheless, the purple protein is a conspicuous presence in the live organisms and it is highly likely that it enhances the bioluminescence of Latia in nature. 

On the basis of the luciferin-luciferase reaction discovered by Dubois (1887), Michelson, Henry and their associates studied the biochemistry of the Pholas bioluminescence for several years beginning in 1970 (Michelson, 1978). They isolated, purified, and characterized the luciferin and the luciferase, and published about a dozen papers in which the luciferin isolated was referred to as Pholas luciferin. Since the luciferin is clearly a protein, later authors called it pholasin following the traditional way of naming a photoprotein... 

Harvey (1917) noted that the fresh arm photophores of Watasenia scintillans do not show a luciferin-luciferase reaction, and Shoji (1919) using a gas chamber of purified hydrogen demonstrated that molecular oxygen is needed for the luminescence. 

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. 7.2.8 Influence of pH on the luminescence intensity of the Odontosyllis luciferin-luciferase reaction at room temperature. From Shimomura et ai, 1963d, with permission from John Wiley Sc Sons Ltd.

Regarding the polychaete Tomopteris, the author has briefly examined its bioluminescence using a few specimens donated by Dr. Steven Haddock. The specimens did not show a luciferin-luciferase reaction, as noted by Harvey (1952). The luminescence could be... 

Fig. 8.1 Luminescence spectra of luciferin-luciferase reaction of the dinoflagellate Gonyaulax polyedra (Lingulodinium polyedrum) in a solution (solid line), isolated scintillons (x), and living Gonyaulax cells (o). From Hastings et al., 1966.

Luciferin binding protein (LBP) binds luciferin at pH 8 but not at pH 6 (Fogel and Hastings, 1971) thus, LBP inhibits the luciferin-luciferase reaction at pH 8 but not at pH 6. Luciferin bound to LBP is stable, differing from the free form of luciferin that is extremely unstable. The molecular size of the Gonyaulax LBP was considered... 

Efforts by various investigators to demonstrate a luciferin-luciferase reaction with luminous fungi were uniformly unsuccessful until Airth and McElroy (1959) obtained a positive result. Airth s group studied further details of the luciferin-luciferase reaction using the luciferin preparation obtained from Armillaria mellea and the luciferase preparation obtained from a species identified by them... 

The preparations of luciferin (Ln, an electron acceptor) and soluble enzyme used were crude or only partially purified. The luciferase was an insoluble particulate material, possibly composed of many substances having various functions. Moreover, the luciferin-luciferase reaction was negative when both luciferin and luciferase were prepared from certain species of luminous fungus. It appears that the light production reported was the result of a complex mechanism involving unknown substances in the test mixture, and probably the crucial step of the light-emitting reaction is not represented by the above schemes. 

Chemiluminescent compounds and their precursors in P. stipticus. Although P. stipticus is negative in the luciferin-luciferase reaction, crude extracts of this fungus are chemiluminescent, like the luciferin obtained from Ompbalia flavida by Kuwabara and Wassink (1966). The chemiluminescence is elicited by the addition of H2O2 and Fe2+ under a mild condition of pH 5-8, and the luminescence is strongly... 

The role of a cationic surfactant must be to provide a necessary hydrophobic and polarized environment for the molecule of luciferin for its luminescence reaction. In the case of a common luciferin-luciferase reaction, such an environment is provided by the enzyme luciferase. The chemical structures of PMs, as well as that of the natural luciferin, have not been determined yet (see the next section). 

Dure and Cormier (1961) demonstrated a luciferin-luciferase reaction for the first time in the extracts of the acorn worm Balanoglossus biminiensis, and also discovered that the luminescence reaction is stimulated by H2O2, of which the details are described below. Recently, Kanakubo and Isobe (2005) reported the chemical structure of a probable luciferin of another acorn worm Ptychodera flava. 

According to Dure and Cormier (1961, 1963) and Cormier and Dure (1963), they made the preparations of luciferase and luciferin from Balanoglossus biminiensis, collected on Sapelo Island, Georgia, and investigated the luciferin-luciferase reaction, as summarized below. 

Haneda, Y., and Johnson, F. H. (1958). The luciferin-luciferase reaction in a fish, Parapriacanthus beryciformis, of newly discovered luminescence. Proc. Natl. Acad. Sci. USA 44 127-129. 

Strehler, B. L., and Cormier, M. J. (1954). Isolation, identification, and function of long chain fatty aldehydes affecting the bacterial luciferin-luciferase reaction. J. Biol. Chem. 211 213-225. 

Wood, K. V. (1993a). Luciferin-luciferase reaction in luminous flies. Bioluminescence Symposium abstract, Nov. 5-10, 1993, Kaanapali Beach, Hawaii, p. 64. 

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