Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.14.14.3 (luciferase)
 38,195 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

2,4-Dinitrofluorobenzene (DNFB) changes the parameters of the bioluminescent reaction involving two enzymes, i.e. NADH: FMN-oxidoreductase and luciferase, by decreasing the maximal intensity of luminescence and increasing the time of maximal intensity. Modification of the proteins does not affect the changes of the reaction parameters. The effects of DNFB on each one of the reactions, i.e. reduction of FNM and light emission, were studied. DNFB does not inhibit the reaction of FMN reduction. It was shown that the mechanism of DNFB effect of bioluminescence consists in uncoupling of FMN reduction and light emission due to competitive inhibition of the FMNH2 active center by 2,4-dinitrofluorobenzene.
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PMID:[Mechanism of action of 2,4-dinitrofluorobenzene on bacterial luminescence in vitro]. 721 55

A procedure resulting in a highly purified preparation of bacterial luciferase with a high specific activity towards FMNH2 and NADH was developed. Using SDS-electrophoresis in polyacrylamide gel, it was shown that the enzyme has a subunit composition and that its monomers do not contain the luciferase activity. The use of the obtained preparation for determining the content of NADH and FMN by the bioluminescent method allowed to increase its sensitivity up to 10(-18) and 10(-17) moles, respectively.
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PMID:[Isolation and purification of bacterial luciferase from Photobacterium fischeri for analytical purposes]. 724 58

The quenching of luminescence of bacterial luciferase from Photobacterium fischeri by non-specific electron acceptors and inhibitors of dehydrogenases was studied. The inhibition of the luminescent reaction obeys the non-competitive mechanism with NADH, FMN and aliphatic aldehyde. The inhibitors compete with cytochrome c for NADH -- cytochrome c oxido-reductase. It is concluded that lumiredoxin, a FeS-containing protein, is the most sensitive component of the luminescent electron transport chain.
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PMID:[Inhibitory analysis of the luminescent electron transport chain of Photobacterium fischeri]. 724 74

Fatty acid reductase from the bioluminescent bacterium Photobacterium phosphoreum, has been partially purified free of aldehyde reductase activity and with a low endogenous fatty acid content permitting the characterization of the aldehyde product of the reaction. Two aldehyde reductases, both dependent on NADH, were separated by anion-exchange chromatography from the fatty acid reductase activity. The partially purified fatty acid reductase catalyzed the synthesis exclusively of long chain aldehydes from fatty acids in the presence of ATP and NADPH as demonstrated by the conversion of [3H]tetradecanoic acid to [3H]aldehyde. Comparison of the amount of [3H]aldehyde produced with the bioluminescence responses of luciferase to the aldehyde product and standard aldehydes, both with respect to maximum light intensity and luminescent decay, established that tetradecanoic acid has been converted to tetradecanal, the aldehyde of the same chain length. These results are consistent with a mechanism involving activation of the fatty acid with ATP followed by reduction of a fatty acyl intermediate to the corresponding aldehyde.
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PMID:Fatty acid reductase in bioluminescent bacteria. Resolution from aldehyde reductases and characterization of the aldehyde product. 729 39

Bioluminescence photokinetic assay of NADP+ is described, using the glucose-6-phosphate dehydrogenase reaction for conversion to its reduced form and subsequent measurement of this with luciferase extracts of Vibria fisherii. the analyses were applied to the determination of the activity of minute amounts of glutathione reductase using NADP+ as measurable product and for nucleotide assay in cell samples of 0.5--10 microgram dry weight. The sensitivity was sufficient for determining 0.5 picomoles NADP+. Previously, FMN, NADH, NAD+ and NADH have been analysed with the bacterial luciferase system. Its applicability has not been extended by the assay of NADP+.
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PMID:Photokinetic microanalysis of NADP+, using bacterial luciferase. 744 56

Particulate fractions of Thiobacillus denitrificans catalyse that the phosphorylation of ADP to ATP during the oxidation of various inorganic sulphur compounds or NADH via an electron transport chain. On the other hand, a "soluble" cell-free fraction synthesized ATP from APS and inorganic phosphate. The production of ATP was verified either by the firefly luciferin-luciferase enzyme system or by the incorporation of 32Pi into ATP. During the oxidation of sulphide, sulphite and NADH the production of ATP from ADP by particulate fractions is inhibited by compounds that inhibit electron transfer and by uncouplers of oxidative phosphorylation. However, these compounds had little effect on the production of ATP from AMP during the oxidation of sulphite by the soluble fraction. NADH was the most effective electron donor for oxidative phosphorylation. The soluble fraction contained high activities of ATP sulphurylase, inorganic pyrophosphatase and adenylate kinase but ADP sulphurylase activity was relatively low. The effects of inhibitors on ATP production from APS and Pi are compared with those on adenylate kinase and ATP sulphurylase.
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PMID:Substrate level versus oxidative phosphorylation in the generation of ATP in Thiobacillus denitrificans. 745 35

Methods that use bacterial luciferase for the assay of NADH in the range from 1 pmol to 1 nmol are described. Optimal conditions for the assay of glycolytic intermediates, tricarboxylic acid-cycle intermediates and related amino acids from milligram amounts of tissue are presented. The whole spectrum of these intermediates can be determined on about 10 mg of liver tissue. The methods are simple, are suitable for routine use, and the instrumentation is inexpensive. The concentrations of glycolytic intermediates in rat livers were determined by conventional spectrometric methods and with luciferase, and the results found to be in good agreement.
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PMID:The determination of reduced nicotinamide-adenine dinucleotide and metabolic intermediates in picomole amounts with bacterial luciferase. 747 36

A fiber optic sensor based on enzyme-catalyzed light-emitting reactions has been developed and integrated in a flow-injection analysis (FIA) system. The firefly luciferase, specific for ATP, and the bacterial oxidoreductase/luciferase system, specific for NADH, have been immobilized on preactivated polyamide membranes. ATP and NADH analysis could be performed in the range from 0.1 pmol to 3 nmol and from 0.5 pmol to 1 nmol, respectively. By co-immobilizing these two bioluminescence systems on the same membrane, a multi-function biosensor has been designed allowing the alternate determination of ATP or NADH with the same sensitivity as that obtained with the two different mono-functional biosensors. A partly self-contained biosensor has been also developed for the flow injection analysis of NADH. For this purpose, FMN (one of the substrates of the bacterial bienzymatic system) has been embedded in a synthetic matrix. Different supports have been tested for the non-covalent immobilization of this substrate and its release in the immediate vicinity of the bound enzymes. Using a photo-crosslinked poly(vinyl alcohol) support, 40 reliable assays (CV = 4.5%) could be performed without changing or reloading the matrix.
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PMID:Design of bioluminescence-based fiber optic sensors for flow-injection analysis. 776 43

The NAD(P)H-flavin oxidoreductase gene from the bioluminescent bacterium, Vibrio fischeri ATCC 7744, was expressed in Escherichia coli, and the enzyme purified using Cibacron Blue 3G-A affinity column chromatography from crude extracts in a single step. The purified enzyme had a typical flavoprotein absorption spectrum and flavin mononucleotide (FMN) was identified as a prosthetic group, non-covalently bound in a molar ratio of 1:1. The enzyme catalyzed the electron transfer from NADH via FMNH2 to various other electron acceptors. Reduced flavin produced by flavin reductase participated non-enzymatically in the following reactions: H2O2-forming NADH oxidase-like, oxygen-insensitive nitroreductase-like, diaphorase (quinone reductase)-like and bacterial luciferase reactions.
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PMID:NAD(P)H-flavin oxidoreductase from the bioluminescent bacterium, Vibrio fischeri ATCC 7744, is a flavoprotein. 803 96

P-flavin-bound luciferase, P-flavin-free luciferase, and P-flavin-bound beta-subunit of luciferase were prepared from Photobacterium phosphoreum using hydrophobic interaction chromatography after conventional purification using DEAE-cellulose chromatography and gel-filtration. The P-flavin-bound luciferase preparation contained about 20% P-flavin-free luciferase not removable by the present procedure. Since the specific activity of the P-flavin-bound luciferase preparation was about 20% of that of the P-flavin-free luciferase, it was concluded that the P-flavin-bound luciferase is an enzyme-product complex and has no more luciferase activity. Unlike the absorption spectrum of FP390 or other flavoproteins, that of P-flavin-bound luciferase preparation has a high absorption peak around 370 nm and resembles the spectrum synthesized by superposing the P-flavin-free luciferase spectrum on the P-flavin-bound beta-subunit spectrum: the P-flavin-bound beta-subunit spectrum is similar to that of FP390, while that of P-flavin-free luciferase has an absorption peak around 370 nm but practically no peak around 450 nm. In addition, P-flavin-free luciferase exhibits a weak but distinct NADH-FMN oxidoreductase activity. These results suggest that a prosthetic group, which absorbs around 370 nm, binds to the luciferase and that this compound is required to yield P-flavin; and they support the hypothesis that the physiological function of bacterial luciferase is to produce P-flavin. Furthermore, the presence of P-flavin-bound beta-subunit of the luciferase in the cell extract supports the hypothesis that physiological function of the lux operon is the biosynthesis of FP390 including its prosthetic group.
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PMID:Preparation of P-flavin-bound and P-flavin-free luciferase and P-flavin-bound beta-subunit of luciferase from Photobacterium phosphoreum. 808 82


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