Firefly luciferin

Last updated
Firefly luciferin
Firefly luciferin.svg
Names
IUPAC name
(4S)-2-(6-hydroxy-1,3-benzothiazol-2-yl)-4,5-dihydrothiazole-4-carboxylic acid
Other names
D-(−)-Luciferin, beetle luciferin
Identifiers
3D model (JSmol)
ChemSpider
ECHA InfoCard 100.018.166 OOjs UI icon edit-ltr-progressive.svg
EC Number
  • 219-981-3
PubChem CID
UNII
  • InChI=1S/C11H8N2O3S2/c14-5-1-2-6-8(3-5)18-10(12-6)9-13-7(4-17-9)11(15)16/h1-3,7,14H,4H2,(H,15,16)/t7-/m1/s1
    Key: BJGNCJDXODQBOB-SSDOTTSWSA-N
  • One of the other tautomeric representations:InChI=1S/C11H8N2O3S2/c14-5-1-2-6-8(3-5)18-10(12-6)9-13-7(4-17-9)11(15)16/h1-3,7,13H,4H2,(H,15,16)/b10-9+/t7-/m1/s1
  • InChI=1S/C11H8N2O3S2/c14-5-1-2-6-8(3-5)18-10(12-6)9-13-7(4-17-9)11(15)16/h1-3,7,14H,4H2,(H,15,16)/t7-/m1/s1
  • O=C(O)[C@@H]1/N=C(\SC1)c2sc3cc(O)ccc3n2
Properties
C11H8N2O3S2
Molar mass 280.32 g·mol−1
UV-vismax)330 nm (neutral and somewhat acidic aqueous solutions) [1]
Absorbance ε330 = 18.2 mM−1 cm−1 [1]
Hazards
GHS labelling:
GHS-pictogram-exclam.svg
Warning
H315, H319, H335
P261, P264, P271, P280, P302+P352, P304+P340, P305+P351+P338, P312, P321, P332+P313, P337+P313, P362, P403+P233, P405, P501
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

Firefly luciferin (also known as beetle luciferin) is the luciferin, or light-emitting compound, used for the firefly (Lampyridae), railroad worm (Phengodidae), starworm (Rhagophthalmidae), and click-beetle (Pyrophorini) bioluminescent systems. It is the substrate of luciferase (EC 1.13.12.7), which is responsible for the characteristic yellow light emission from many firefly species.

Contents

As with all other luciferins, oxygen is required to elicit light; however, it has also been found adenosine triphosphate (ATP) and magnesium are required for light emission. [2] [3]

History

Much of the early work on the chemistry of the firefly luminescence was done in the lab of William D. McElroy at Johns Hopkins University. The luciferin was first isolated and purified in 1949, though it would be several years until a procedure was developed to crystallize the compound in high yield. This, along with the synthesis and structure elucidation, was accomplished by Dr. Emil H. White at the Johns Hopkins University, Department of Chemistry. [4] The procedure was an acid-base extraction, given the carboxylic acid group on the luciferin. The luciferin could be effectively extracted using ethyl acetate at low pH from powder of approximately 15,000 firefly lanterns. [5] The structure was later confirmed by combined use of infrared spectroscopy, UV–vis spectroscopy and synthetic methods to degrade the compound into identifiable fragments. [6]

Properties

Crystal luciferin was found to be fluorescent, absorbing ultraviolet light with a peak at 327 nm and emitting light with a peak at 530 nm. Visible emission occurs upon relaxation of the oxyluciferin from a singlet excited state down to its ground state. [7] Alkaline solutions caused a redshift of the absorption likely due to deprotonation of the hydroxyl group on the benzothiazole, but did not affect the fluorescence emission. It was found that the luciferyl adenylate (the AMP ester of luciferin) spontaneously emits light in solution. [8] Different species of fireflies all use the same luciferin, however the color of the light emitted can differ greatly. The light from Photuris pennsylvanica was measured to be 552 nm (green-yellow) while Pyrophorus plagiophthalamus was measured to emit light at 582 nm (orange) in the ventral organ. Such differences are likely due to pH changes or differences in primary structure of the luciferase. [9] Modification of the firefly luciferin substrate has led to "red-shifted" emissions (up to emission wavelength of 675 nm). [10]

Biological activity

The in vivo synthesis of firefly luciferin is not completely understood. Only the final step of the enzymatic pathway has been studied, which is the condensation reaction of D-cysteine with 2-cyano-6-hydroxybenzothiazole, and is the same reaction used to produce the compound synthetically. [11] This was confirmed by radiolabeling of atoms in the two compounds and by identification of a luciferin-regenerating enzyme. [12]

In firefly, oxidation of luciferins, which is catalyzed by luciferases, yields a peroxy compound 1,2-dioxetane. The dioxetane is unstable and decays spontaneously to carbon dioxide and excited ketones, which release excess energy by emitting light (bioluminescence). [13]

Loss of CO2 of a dioxetane, giving rise to an excited ketone, which relaxes by emitting light. Luciferin principle.png
Loss of CO2 of a dioxetane, giving rise to an excited ketone, which relaxes by emitting light.

Firefly luciferin and modified substrates are fatty acid mimics and have been used to localize fatty acid amide hydrolase (FAAH) in vivo. [14] Firefly luciferin is a substrate of the ABCG2 transporter and has been used as part of a bioluminescence imaging high throughput assay to screen for inhibitors of the transporter. [15]

Related Research Articles

<span class="mw-page-title-main">Bioluminescence</span> Emission of light by a living organism

Bioluminescence is the production and emission of light by living organisms. It is a form of chemiluminescence. Bioluminescence occurs widely in marine vertebrates and invertebrates, as well as in some fungi, microorganisms including some bioluminescent bacteria, and terrestrial arthropods such as fireflies. In some animals, the light is bacteriogenic, produced by symbiotic bacteria such as those from the genus Vibrio; in others, it is autogenic, produced by the animals themselves.

<span class="mw-page-title-main">Chemiluminescence</span> Emission of light as a result of a chemical reaction

Chemiluminescence is the emission of light (luminescence) as the result of a chemical reaction, i.e. a chemical reaction results in a flash or glow of light. A standard example of chemiluminescence in the laboratory setting is the luminol test. Here, blood is indicated by luminescence upon contact with iron in hemoglobin. When chemiluminescence takes place in living organisms, the phenomenon is called bioluminescence. A light stick emits light by chemiluminescence.

<span class="mw-page-title-main">Luciferase</span> Enzyme family

Luciferase is a generic term for the class of oxidative enzymes that produce bioluminescence, and is usually distinguished from a photoprotein. The name was first used by Raphaël Dubois who invented the words luciferin and luciferase, for the substrate and enzyme, respectively. Both words are derived from the Latin word lucifer, meaning "lightbearer", which in turn is derived from the Latin words for "light" (lux) and "to bring or carry" (ferre).

<span class="mw-page-title-main">Luciferin</span> Class of light-emitting chemical compounds

Luciferin is a generic term for the light-emitting compound found in organisms that generate bioluminescence. Luciferins typically undergo an enzyme-catalyzed reaction with molecular oxygen. The resulting transformation, which usually involves breaking off a molecular fragment, produces an excited state intermediate that emits light upon decaying to its ground state. The term may refer to molecules that are substrates for both luciferases and photoproteins.

<span class="mw-page-title-main">Organic peroxides</span> Organic compounds of the form R–O–O–R’

In organic chemistry, organic peroxides are organic compounds containing the peroxide functional group. If the R′ is hydrogen, the compounds are called hydroperoxides, which are discussed in that article. The O−O bond of peroxides easily breaks, producing free radicals of the form RO. Thus, organic peroxides are useful as initiators for some types of polymerization, such as the acrylic, unsaturated polyester, and vinyl ester resins used in glass-reinforced plastics. MEKP and benzoyl peroxide are commonly used for this purpose. However, the same property also means that organic peroxides can explosively combust. Organic peroxides, like their inorganic counterparts, are often powerful bleaching agents.

<span class="mw-page-title-main">Firefly luciferase</span>

Firefly luciferase is the light-emitting enzyme responsible for the bioluminescence of fireflies and click beetles. The enzyme catalyses the oxidation of firefly luciferin, requiring oxygen and ATP. Because of the requirement of ATP, firefly luciferases have been used extensively in biotechnology.

<span class="mw-page-title-main">Bioluminescence imaging</span>

Bioluminescence imaging (BLI) is a technology developed over the past decades (1990's and onward). that allows for the noninvasive study of ongoing biological processes Recently, bioluminescence tomography (BLT) has become possible and several systems have become commercially available. In 2011, PerkinElmer acquired one of the most popular lines of optical imaging systems with bioluminescence from Caliper Life Sciences.

A photocyte is a cell that specializes in catalyzing enzymes to produce light (bioluminescence). Photocytes typically occur in select layers of epithelial tissue, functioning singly or in a group, or as part of a larger apparatus. They contain special structures called photocyte granules. These specialized cells are found in a range of multicellular animals including ctenophora, coelenterates (cnidaria), annelids, arthropoda and fishes. Although some fungi are bioluminescent, they do not have such specialized cells.

The chemical substance 1,2-dioxetane is a heterocyclic, organic compound with formula C2O2H4, containing a ring of two adjacent oxygen atoms and two adjacent carbon atoms. It is therefore an organic peroxide, and can be viewed as a dimer of formaldehyde.

In enzymology, an Oplophorus-luciferin 2-monooxygenase, also known as Oplophorus luciferase is a luciferase, an enzyme, from the deep-sea shrimp Oplophorus gracilirostris [2], belonging to a group of coelenterazine luciferases. Unlike other luciferases, it has a broader substrate specificity [3,4,6] and can also bind to bisdeoxycoelenterazine efficiently [3,4]. It is the third example of a luciferase to be purified in lab [2]. The systematic name of this enzyme class is Oplophorus-luciferin:oxygen 2-oxidoreductase (decarboxylating). This enzyme is also called Oplophorus luciferase.

<span class="mw-page-title-main">Renilla-luciferin 2-monooxygenase</span>

Renilla-luciferin 2-monooxygenase, Renilla luciferase, or RLuc, is a bioluminescent enzyme found in Renilla reniformis, belonging to a group of coelenterazine luciferases. Of this group of enzymes, the luciferase from Renilla reniformis has been the most extensively studied, and due to its bioluminescence requiring only molecular oxygen, has a wide range of applications, with uses as a reporter gene probe in cell culture, in vivo imaging, and various other areas of biological research. Recently, chimeras of RLuc have been developed and demonstrated to be the brightest luminescent proteins to date, and have proved effective in both noninvasive single-cell and whole body imaging.

<span class="mw-page-title-main">Bioreporter</span> Genetically engineered microbial cells

Bioreporters are intact, living microbial cells that have been genetically engineered to produce a measurable signal in response to a specific chemical or physical agent in their environment. Bioreporters contain two essential genetic elements, a promoter gene and a reporter gene. The promoter gene is turned on (transcribed) when the target agent is present in the cell’s environment. The promoter gene in a normal bacterial cell is linked to other genes that are then likewise transcribed and then translated into proteins that help the cell in either combating or adapting to the agent to which it has been exposed. In the case of a bioreporter, these genes, or portions thereof, have been removed and replaced with a reporter gene. As a result, turning on the promoter gene also turns on the reporter gene, leading to the production of reporter proteins that output a detectable signal. The presence of a signal indicates that the bioreporter has sensed a particular agent in its environment.

<span class="mw-page-title-main">Coelenterazine</span> Chemical compound

Coelenterazine is a luciferin, a molecule that emits light after reaction with oxygen, found in many aquatic organisms across eight phyla. It is the substrate of many luciferases such as Renilla reniformis luciferase (Rluc), Gaussia luciferase (Gluc), and photoproteins, including aequorin, and obelin. All these proteins catalyze the oxidation of this substance, a reaction catalogued EC 1.13.12.5.

<span class="mw-page-title-main">Vargulin</span> Chemical compound

Vargulin, also called Cypridinid luciferin, Cypridina luciferin, or Vargula luciferin, is the luciferin found in the ostracod Cypridina hilgendorfii, also named Vargula hilgendorfii. These bottom dwelling ostracods emit a light stream into water when disturbed presumably to deter predation. Vargulin is also used by the midshipman fish, Porichthys.

<i>Vargula hilgendorfii</i> Species of seed shrimp

Vargula hilgendorfii, sometimes called the sea-firefly and one of three bioluminescent species known in Japan as umi-hotaru (海蛍), is a species of ostracod crustacean. It is the only member of genus Vargula to inhabit Japanese waters; all other members of its genus inhabit the Gulf of Mexico, the Caribbean Sea, and waters off the coast of California. V. hilgendorfii was formerly more common, but its numbers have fallen significantly.

<span class="mw-page-title-main">John Woodland Hastings</span>

John Woodland "Woody" Hastings, was a leader in the field of photobiology, especially bioluminescence, and was one of the founders of the field of circadian biology. He was the Paul C. Mangelsdorf Professor of Natural Sciences and Professor of Molecular and Cellular Biology at Harvard University. He published over 400 papers and co-edited three books.

<span class="mw-page-title-main">Dinoflagellate luciferase</span>

Dinoflagellate luciferase (EC 1.13.12.18, Gonyaulax luciferase) is a specific luciferase, an enzyme with systematic name dinoflagellate-luciferin:oxygen 132-oxidoreductase.

<span class="mw-page-title-main">Bioluminescent bacteria</span>

Bioluminescent bacteria are light-producing bacteria that are predominantly present in sea water, marine sediments, the surface of decomposing fish and in the gut of marine animals. While not as common, bacterial bioluminescence is also found in terrestrial and freshwater bacteria. These bacteria may be free living or in symbiosis with animals such as the Hawaiian Bobtail squid or terrestrial nematodes. The host organisms provide these bacteria a safe home and sufficient nutrition. In exchange, the hosts use the light produced by the bacteria for camouflage, prey and/or mate attraction. Bioluminescent bacteria have evolved symbiotic relationships with other organisms in which both participants benefit close to equally. Another possible reason bacteria use luminescence reaction is for quorum sensing, an ability to regulate gene expression in response to bacterial cell density.

<span class="mw-page-title-main">Scintillon</span>

Scintillons are small structures in cytoplasm that produce light. Among bioluminescent organisms, only dinoflagellates have scintillons.

Howard Harold Seliger was a physicist, biochemist, and biology professor, known for his research on bioluminescence.

References

  1. 1 2 "D-luciferin product information" (PDF). Sigma Aldrich.
  2. McElroy WD (1947). "The Energy Source for Bioluminescence in an Isolated System". Proc Natl Acad Sci USA. 33 (11): 342–345. Bibcode:1947PNAS...33..342M. doi: 10.1073/pnas.33.11.342 . PMC   1079070 . PMID   16588763.
  3. Green A, McElroy WD (1956). "Function of adenosine triphosphate in the activation of luciferin". Arch Biochem Biophys. 64 (2): 257–271. doi:10.1016/0003-9861(56)90268-5. PMID   13363432.
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  5. Bitler B, McElroy WD (1957). "The Preparation and Properties of Crystalline Firely Luciferin". Arch Biochem Biophys. 72 (2): 358–368. doi:10.1016/0003-9861(57)90212-6. PMID   13479120.
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  7. Marques SM, Joaquim (2009). "Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions". IUBMB Life. 61 (1): 6–17. doi: 10.1002/iub.134 . PMID   18949818. S2CID   21583225.
  8. Rhodes WC, McElroy WD (1958). "The synthesis and function of luciferyl-adenylate and oxyluciferyl-adenylate". J Biol Chem. 233 (6): 1528–1537. doi: 10.1016/S0021-9258(18)49367-2 . PMID   13610868.
  9. Seliger HH, Buck JB, Fastie WG, McElroy WD (1964). "The Spectral Distribution of Firefly Light". J Gen Physiol. 48 (1): 95–104. doi:10.1085/jgp.48.1.95. PMC   2195396 . PMID   14212153.
  10. Kiyama M, Saito R, Iwano S, Obata R, Niwa H, Maki SA (2016). "Multicolor Bioluminescence Obtained Using Firefly Luciferin". Current Topics in Medicinal Chemistry. 16 (24): 2648–2655. doi:10.2174/1568026616666160413135055. PMID   27072707.
  11. White EH, Worther H, Field GF, McElroy WD (1965). "Analogs of Firefly Luciferin". J. Org. Chem. 30 (7): 2344–2348. doi:10.1021/jo01018a054.
  12. Gomi K, Kajiyama N (2001). "Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin". J Biol Chem. 276 (39): 36508–36513. doi: 10.1074/jbc.M105528200 . PMID   11457857.
  13. Aldo Roda Chemiluminescence and Bioluminescence: Past, Present and Future, p. 57, Royal Society of Chemistry, 2010, ISBN   1-84755-812-7
  14. Mofford DM, Adams ST, Kumar Reddy GS, Randheer Reddy G, Miller SC (2015). "Luciferin Amides Enable in Vivo Bioluminescence Detection of Endogenous Fatty Acid Amide Hydrolase Activity". J. Am. Chem. Soc. 137 (27): 8684–8687. doi:10.1021/jacs.5b04357. PMC   4507478 . PMID   26120870.
  15. "Identification of Inhibitors of ABCG2 by a Bioluminescence Imaging-Based High-Throughput Assay". Cancer Res. 69.