Firefly Luciferase Produces Hydrogen Peroxide as a Coproduct in Dehydroluciferyl Adenylate Formation

Firefly luciferase catalyzes the synthesis of H2O2 from the same substrates as the bioluminescence reaction: ATP and luciferin (d‐LH2). About 80 % of the enzyme‐bound intermediate d‐luciferyl adenylate (d‐LH2‐AMP) is oxidized into oxyluciferin, and a photon is emitted during this reaction. The enzyme pathway responsible for the generation of H2O2 is a side reaction in which d‐LH2‐AMP is oxidized into dehydroluciferyl adenylate (L‐AMP). Like the bioluminescence reaction, the luciferase‐catalyzed synthesis of H2O2 and L‐AMP is a stereospecific process, involving only the natural D enantiomer. However, the intramolecular electron transfer postulated as essential to the light emission process is not involved in this side reaction.

[1]  J. E. D. Esteves da Silva,et al.  Coenzyme A affects firefly luciferase luminescence because it acts as a substrate and not as an allosteric effector , 2005, The FEBS journal.

[2]  T. Hirano,et al.  Firefly luciferase exhibits bimodal action depending on the luciferin chirality. , 2005, Biochemical and biophysical research communications.

[3]  J. E. D. Esteves da Silva,et al.  Synthesis of luciferyl coenzyme A: a bioluminescent substrate for firefly luciferase in the presence of AMP. , 2005, Angewandte Chemie.

[4]  J. E. D. Esteves da Silva,et al.  Identification of Luciferyl Adenylate and Luciferyl Coenzyme A Synthesized by Firefly Luciferase , 2004, Chembiochem : a European journal of chemical biology.

[5]  J. E. D. Esteves da Silva,et al.  pH opposite effects on synthesis of dinucleoside polyphosphates and on oxidation reactions catalyzed by firefly luciferase , 2003, FEBS letters.

[6]  V. Viviani,et al.  The origin, diversity, and structure function relationships of insect luciferases , 2002, Cellular and Molecular Life Sciences CMLS.

[7]  Sanjiv S Gambhir,et al.  Optical Imaging of Cardiac Reporter Gene Expression in Living Rats , 2002, Circulation.

[8]  J. Stroh,et al.  Yellow-green and red firefly bioluminescence from 5,5-dimethyloxyluciferin. , 2002, Journal of the American Chemical Society.

[9]  J. C. D. Silva,et al.  Identification of enzyme produced firefly oxyluciferin by reverse phase HPLC , 2001 .

[10]  Antonio Sillero,et al.  Dehydroluciferyl‐AMP is the main intermediate in the luciferin dependent synthesis of Ap4A catalyzed by firefly luciferase , 1998, FEBS letters.

[11]  R. Haugland,et al.  A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. , 1997, Analytical biochemistry.

[12]  Masafumi Oshiro,et al.  Visualizing Gene Expression in Living Mammals Using a Bioluminescent Reporter , 1997, Photochemistry and photobiology.

[13]  A. Sillero,et al.  Synthesis of dehydroluciferin by firefly luciferase: effect of dehydroluciferin, coenzyme A and nucleoside triphosphates on the luminescent reaction. , 1997, Biochemical and biophysical research communications.

[14]  J. Stroh,et al.  Identification of a Firefly Luciferase Active Site Peptide Using a Benzophenone-based Photooxidation Reagent* , 1997, The Journal of Biological Chemistry.

[15]  E. Schulman,et al.  A highly sensitive fluorescent micro-assay of H2O2 release from activated human leukocytes using a dihydroxyphenoxazine derivative. , 1997, Journal of immunological methods.

[16]  N. Lembert.  Firefly luciferase can use L-luciferin to produce light. , 1996, The Biochemical journal.

[17]  I. Leclercq,et al.  Fast determination of low-level cytochrome P-450 1A1 activity by high-performance liquid chromatography with fluorescence or visible absorbance detection. , 1996, Journal of chromatography. B, Biomedical applications.

[18]  K. Wood.  THE CHEMICAL MECHANISM and EVOLUTIONARY DEVELOPMENT OF BEETLE BIOLUMINESCENCE , 1995 .

[19]  A. Sillero,et al.  Influence of oxygen, dehydroluciferin, luciferin and 6′-ethyl-luciferin on the synthesis of adenosine (5′) tetraphospho (5′) adenosine (Ap4A) by firefly luciferase , 1995 .

[20]  B. Demple,et al.  Intracellular generation of superoxide as a by-product of Vibrio harveyi luciferase expressed in Escherichia coli , 1994, Journal of bacteriology.

[21]  L. Y. Brovko,et al.  Oxyluciferin fluorescence is a model of native bioluminescence in the firefly luciferin—luciferase system , 1993 .

[22]  P. Brennan,et al.  NAPHTHYL‐ AND QUINOLYLLUCIFERIN: GREEN AND RED LIGHT EMITTING FIREFLY LUCIFERIN ANALOGUES , 1989, Photochemistry and photobiology.

[23]  S. Gould,et al.  Firefly luciferase is targeted to peroxisomes in mammalian cells. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[24]  B. Halliwell,et al.  Free radicals in biology and medicine , 1985 .

[25]  R. Holmes,et al.  A spectrophotometric procedure for determining the activity of various rat tissue oxidases. , 1975, Analytical Biochemistry.

[26]  H. Seliger,et al.  The spectroscopic properties of firefly luciferin and related compounds. An approach to product emission. , 1969, Biochemistry.

[27]  Y. Kishi,et al.  Luciferine, bioluminescierende Substanzen , 1968 .

[28]  Y. Kishi,et al.  Luciferins, bioluminescent substances. , 1968, Angewandte Chemie.

[29]  E. H. White,et al.  Analogs of firefly luciferin. 3. , 1965, The Journal of organic chemistry.

[30]  W. D. Mcelroy,et al.  Stereo-specificity and firefly bioluminescence, a comparison of natural and synthetic luciferins. , 1961, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Emil H. White,et al.  THE STRUCTURE AND SYNTHESIS OF FIREFLY LUCIFERIN , 1961 .

[32]  W. D. Mcelroy,et al.  The synthesis and function of luciferyl-adenylate and oxyluciferyl-adenylate. , 1958, The Journal of biological chemistry.

[33]  W. D. Mcelroy,et al.  Function of adenosine triphosphate in the activation of luciferin. , 1956, Archives of biochemistry and biophysics.

[34]  Jianfeng Xu,et al.  Sequence variants in the 3'-->5' deoxyribonuclease TREX2: identification in a genetic screen and effects on catalysis by the recombinant proteins. , 2004, Advances in enzyme regulation.

[35]  J. Hastings,et al.  Bioluminescence and chemiluminescence. , 2003, Methods in enzymology.

[36]  Kevin R. Messner,et al.  In vitro quantitation of biological superoxide and hydrogen peroxide generation. , 2002, Methods in enzymology.

[37]  A. Lundin.  Use of firefly luciferase in ATP-related assays of biomass, enzymes, and metabolites. , 2000, Methods in enzymology.

[38]  N. Vermeulen,et al.  Oxygen and xenobiotic reductase activities of cytochrome P450. , 1995, Critical reviews in toxicology.

[39]  L. Bowie.  [2] Synthesis of firefly luciferin and structural analogs , 1978 .

[40]  G. Schuster,et al.  Bioluminescence of the firefly: key steps in the formation of the electronically excited state for model systems. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[41]  T. Goto,et al.  Firefly bioluminescence II. Identification of 2-(6′-hydroxybenzothiazol-2′-yl)-4-hydroxythiazole as a product in the bioluminescence of firefly lanterns and as a product in the chemiluminescence of firefly luciferin in DMSO , 1971 .

[42]  W. D. Mcelroy,et al.  The function of coenzyme A in luminescence. , 1958, Biochimica et biophysica acta.