Imidazolopyrazine bioluminescence in copepods and other marine organisms

Campbell, Anthony K.; Herring, Peter J.

doi:10.1007/bf01313261

Cited by 96 publications

(70 citation statements)

References 20 publications

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“…To date primary sequences have been determined for several luciferases (Renilla (32), Oplophorus (24), Pleuromamma, and Gaussia luciferases) and for several Ca 2ϩ -regulated photoproteins (33,34), all of which catalyze the luminescent oxidation of the same substrate, coelenterazine (35). Despite the fact that all of them use the same substrate, it is very intriguing and possibly significant to find no sequence similarity among these bioluminescent proteins.…”

Section: Resultsmentioning

confidence: 99%

Cloning and Expression of cDNA for a Luciferase from the Marine Copepod Metridia longa

Markova

Gölz

Frank

et al. 2004

Journal of Biological Chemistry

141

133

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Metridia longa is a marine copepod from which a blue bioluminescence originates as a secretion from epidermal glands in response to various stimuli. We demonstrate that Metridia luciferase is specific for coelenterazine to produce blue light ( max ‫؍‬ 480 nm). Using an expression cDNA library and functional screening, we cloned and sequenced the cDNA encoding the Metridia luciferase. The cDNA is an 897-bp fragment with a 656-bp open reading frame, which encodes a 219-amino acid polypeptide with a molecular weight of 23,885. The polypeptide contains an N-terminal signal peptide of 17 amino acid residues for secretion. On expression of the Metridia luciferase gene in mammalian Chinese hamster ovary cells the luciferase is detected in the culture medium confirming the existence of a naturally occurring signal peptide for secretion in the cloned luciferase. The novel secreted luciferase was tested in a practical assay application in which the activity of A2a and NPY2 G-protein-coupled receptors was detected. These results clearly suggest that the secreted Metridia luciferase is well suited as a reporter for monitoring gene expression and, in particular, for the development of novel ultrahigh throughput screening technologies.Continuous monitoring of dynamic changes in gene expression from living cells in response to various stimuli provides important information about cell physiology. For this purpose bioluminescent and fluorescent reporters have been introduced as tools for sensitive and convenient monitoring of gene expression. A cDNA encoding a bioluminescent or fluorescent reporter such as a luciferase or green fluorescent protein (GFP) 1 is fused to the promoter region of the target gene, and the construct is transfected to mammalian cells. The gene expression is monitored simply by measuring light emitted through an enzymatic reaction or fluorescence. To date several bioluminescent proteins have been widely and successfully used for optical monitoring of gene expression in living cells: firefly luciferase (FL) (for review, see Refs. 1-3), bacterial luciferase (for review, see Refs. 4 -6), the Ca 2ϩ -regulated photoprotein aequorin (for review, see Refs. 7-9), Renilla luciferase (for review, see Refs. 10 and 11), and GFP (for review, see . Although time course studies of gene expression have been determined by real time imaging with these reporter genes, intracellular assays present difficulties for continuous measurements in some cases. For instance, it is not easy to keep an intracellular concentration of luciferin at a constant level in the case of application of firefly and Renilla luciferases. In the case of FL, additional methods may be required to facilitate the passage of the substrate across the cell membrane. In addition, the intensity and stability of the bioluminescent response of FL in living cells are reported to be affected by concentrations of ATP, luciferin, and luciferin-luciferase complex (15, 16). Despite the fact that the GFP possesses excellent properties as a reporter, it has some short...

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Section: Resultsmentioning

confidence: 99%

Cloning and Expression of cDNA for a Luciferase from the Marine Copepod Metridia longa

Markova

Gölz

Frank

et al. 2004

Journal of Biological Chemistry

141

133

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“…In addition, coelenterazine, the imidazolopyrazine luciferin of these photoprotein systems, is also used by fish, squid, some crustaceans, and a chaetognath (13)(14)(15) and is found in many nonluminous organisms as well (16,17). Despite its occurrence in a variety of phyla, and recent interest in its antioxidative properties (18,19), there has been little experimental evidence to indicate the origins of this light-emitting molecule in nature.…”

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confidence: 99%

Can coelenterates make coelenterazine? Dietary requirement for luciferin in cnidarian bioluminescence

Haddock

Rivers²,

Robison³

2001

Proc. Natl. Acad. Sci. U.S.A.

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In the calcium-activated photoprotein aequorin, light is produced by the oxidation of coelenterazine, the luciferin used by at least seven marine phyla. However, despite extensive research on photoproteins, there has been no evidence to indicate the origin of coelenterazine within the phylum Cnidaria. Here we report that the hydromedusa Aequorea victoria is unable to produce its own coelenterazine and is dependent on a dietary supply of this luciferin for bioluminescence. Although they contain functional apophotoproteins, medusae reared on a luciferin-free diet are unable to produce light unless provided with coelenterazine from an external source. This evidence regarding the origins of luciferin in Cnidaria has implications for the evolution of bioluminescence and for the extensive use of coelenterazine among marine organisms.

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“…Obelia geniculata was collected in Plymouth Sound, Devon, UK, during JuneSeptember 1986JuneSeptember -1989. Copepods and radiolarians were collected on RRS Discovery Cruises 105, 120 and 168 in the eastern North Atlantic (Campbell and Herring, 1989).…”

Section: Organismsmentioning

confidence: 99%

“…In contrast imidazolopyrazine bioluminescence occurs in marine luminous organisms representing six phyla (Saromastigophora/Radiolaria; Cnidaria; Ctenophora; Arthropoda; Mollusca; Chordata) (Campbell and Herring, 1989). Synthesis of luciferase or photoprotein in these organisms can be confirmed by translation of crude RNA or purified followed by addition of benzothiazole in the American firefly and European glow-worm, imidazolopyrazine in the cnidarians Obelia and Aequorea, the radiolarian 7halassicolla, and the copepod Euaugaptilus magnus.…”

Section: /N Vitro Translation Of Mrnamentioning

confidence: 99%

From Luc and Phot genes to the hospital bed

Campbell

Sala-Newby

Aston

et al. 1990

J. Biolumin. Chemilumin.

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The Luc gene from the firefly Photinuspyralis has been isolated by cloning it in pcDVl PL plasmid primer and Honjo linker and the Phot gene isolated from Aequorea victoria using the polymerase chain reaction. A methad has been established using SP6 RNA polymerase for transcribing and translating bioluminescent genes in vitro. It should now be possible t o engineer these genes t o measure intracellular ATP and the covalent modification of proteins in single, live cells, providing unique insights into the molecular basis of disease.

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Imidazolopyrazine bioluminescence in copepods and other marine organisms

Cited by 96 publications

References 20 publications

Cloning and Expression of cDNA for a Luciferase from the Marine Copepod Metridia longa

Cloning and Expression of cDNA for a Luciferase from the Marine Copepod Metridia longa

Can coelenterates make coelenterazine? Dietary requirement for luciferin in cnidarian bioluminescence

From Luc and Phot genes to the hospital bed

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