Interaction of Bacterial Luciferase with 8-Substituted Flavin Mononucleotide Derivatives

Francisco, Wilson A.; Abu-Soûd, Husam M.; Topgi, Ravindra Satish; Baldwin, Thomas O.; Raushel, Frank M.

doi:10.1074/jbc.271.1.104

Cited by 24 publications

(9 citation statements)

References 30 publications

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“…The most commonly exploited luciferases are the terrestrial and marine bacterial luciferases and the eukaryotic firefly and Renilla (Sea Pansy) luciferases (Table 2). Luciferases catalyze the oxidation of reduced flavin mononucleotide (FMNH 2 ) and a long‐chain aliphatic aldehyde in the presence of O 2 to yield blue light (Scheme ) 154. 155 Because FMNH 2 is rapidly oxidized in air, this luciferase cannot provide continuous emission, but instead generates only short bursts of light 137.…”

Section: Biological Materialsmentioning

confidence: 99%

Materials for Fluorescence Resonance Energy Transfer Analysis: Beyond Traditional Donor–Acceptor Combinations

Berti²,

2006

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The use of Förster or fluorescence resonance energy transfer (FRET) as a spectroscopic technique has been in practice for over 50 years. A search of ISI Web of Science with just the acronym "FRET" returns more than 2300 citations from various areas such as structural elucidation of biological molecules and their interactions, in vitro assays, in vivo monitoring in cellular research, nucleic acid analysis, signal transduction, light harvesting and metallic nanomaterials. The advent of new classes of fluorophores including nanocrystals, nanoparticles, polymers, and genetically encoded proteins, in conjunction with ever more sophisticated equipment, has been vital in this development. This review gives a critical overview of the major classes of fluorophore materials that may act as donor, acceptor, or both in a FRET configuration. We focus in particular on the benefits and limitations of these materials and their combinations, as well as the available methods of bioconjugation.

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Section: Biological Materialsmentioning

confidence: 99%

Materials for Fluorescence Resonance Energy Transfer Analysis: Beyond Traditional Donor–Acceptor Combinations

Berti²,

2006

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“…The enzyme responsible for NO production in neurons is the neuronal nitric oxide synthase (nNOS) that mediates the generation of l -citrulline from l -arginine (Figure 3D; Zhou and Zhu, 2009). nNOS responds to oxygen levels and is upregulated by hypoxia in different neurons (AbuSoud et al, 1996; Prabhakar et al, 1996). NO regulates a variety of stress-related transcription factors such as HIF-1 and NF-κB (Contestabile, 2008; Keswani et al, 2011) and can lead to neuronal death under different stress conditions (Brown, 2010).…”

Section: Key Molecules Involvedmentioning

confidence: 99%

Neuronal Responses to Physiological Stress

Kagias¹,

Nehammer²,

Pocock³

2012

Front. Gene.

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Physiological stress can be defined as any external or internal condition that challenges the homeostasis of a cell or an organism. It can be divided into three different aspects: environmental stress, intrinsic developmental stress, and aging. Throughout life all living organisms are challenged by changes in the environment. Fluctuations in oxygen levels, temperature, and redox state for example, trigger molecular events that enable an organism to adapt, survive, and reproduce. In addition to external stressors, organisms experience stress associated with morphogenesis and changes in inner chemistry during normal development. For example, conditions such as intrinsic hypoxia and oxidative stress, due to an increase in tissue mass, have to be confronted by developing embryos in order to complete their development. Finally, organisms face the challenge of stochastic accumulation of molecular damage during aging that results in decline and eventual death. Studies have shown that the nervous system plays a pivotal role in responding to stress. Neurons not only receive and process information from the environment but also actively respond to various stresses to promote survival. These responses include changes in the expression of molecules such as transcription factors and microRNAs that regulate stress resistance and adaptation. Moreover, both intrinsic and extrinsic stresses have a tremendous impact on neuronal development and maintenance with implications in many diseases. Here, we review the responses of neurons to various physiological stressors at the molecular and cellular level.

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“…The second assay method gives a specific activity value approximately double that of the FMNH 2 injection method. Aldehyde binding to free enzyme prevents binding of FMNH 2 , leading to inhibition (45,46). The proposed kinetic mechanism dictates ordered sequential binding of reduced flavin prior to aldehyde (47).…”

Section: Table 2 Kinetic Parameters Obtained From the Fre-luciferase mentioning

confidence: 99%

Fre Is the Major Flavin Reductase Supporting Bioluminescence from Vibrio harveyi Luciferase in Escherichia coli

Campbell

Baldwin

2009

Journal of Biological Chemistry

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Unlike the vast majority of flavoenzymes, bacterial luciferase requires an exogenous source of reduced flavin mononucleotide for bioluminescence activity. Within bioluminescent bacterial cells, species-specific oxidoreductases are believed to provide reduced flavin for luciferase activity. The source of reduced flavin in Escherichia coli-expressing bioluminescence is not known. There are two candidate proteins potentially involved in this process in E. coli, a homolog of the Vibrio harveyi Frp oxidoreductase, NfsA, and a luxG type oxidoreductase, Fre. Using single gene knock-out strains, we show that deletion of fre decreased light output by greater than two orders of magnitude, yet had no effect on luciferase expression in E. coli. Purified Fre is capable of supporting bioluminescence in vitro with activity comparable to that with the endogenous V. harveyi reductase (Frp), using either FMN or riboflavin as substrate. In a pulldown experiment, we found that neither Fre nor Frp co-purify with luciferase. In contrast to prior work, we find no evidence for stable complex formation between luciferase and oxidoreductase. We conclude that in E. coli, an enzyme primarily responsible for riboflavin reduction (Fre) can also be utilized to support high levels of bioluminescence.

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Interaction of Bacterial Luciferase with 8-Substituted Flavin Mononucleotide Derivatives

Cited by 24 publications

References 30 publications

Materials for Fluorescence Resonance Energy Transfer Analysis: Beyond Traditional Donor–Acceptor Combinations

Materials for Fluorescence Resonance Energy Transfer Analysis: Beyond Traditional Donor–Acceptor Combinations

Neuronal Responses to Physiological Stress

Fre Is the Major Flavin Reductase Supporting Bioluminescence from Vibrio harveyi Luciferase in Escherichia coli

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