Bacterial luciferase activators

Eberhard, A. (1972). Inhibition and activation of bacterial luciferase synthesis. J. Bacteriol. 109 1101-1105. 

Escher, A., O Kane, D. J., Lee, J., and Szalay, A. A. (1989). Bacterial luciferase a(3 fusion protein is fully active as a monomer and highly sensitive in vivo to elevated temperature. Proc. Natl. Acad. Sci. USA 86 6528-6532. 

Sinclair, J. F., Waddle, J. J., Waddill, E. F., and Baldwin, T. O. (1993). Purified native subunits of bacterial luciferase are active in the bioluminescence reaction but fail to assemble into the a(3 structure. Biochemistry 32 5036-5044. 

Chemical immobilization procedures of bioluminescent enzymes such as firefly luciferase and bacterial luciferase-NAD(P)H FMN oxidoreductase to glass beads or rods [174, 175], sepharose particles [176], and cellophane films [177] have produced active immobilized enzymes. Picomole-femtomole amounts of ATP or NAD(P)H could be detected using immobilized firefly luciferase or bacterial luciferase-oxidoreductase, respectively. 

Other applications dealt with the development of a luciferin ester substrate to measure the luciferase activity in living cells [141], the detection of toxic compounds such as sodium azide, fluoroacetic acid, and antibiotics [142], the development of a biosensor for the determination of bioavailable mercury [143], the use of eukaryotic luciferases as bacterial markers with different colors of luminescence [144], the determination of complement-mediated killing of bacteria [145], and the development of a bioassay for the determination of HIV type 1 virus and HIV-1 Tat protein activity, valuable also for analysis of HlV-inhibi-tory agents [146],... 

Methods based on chemiluminescent and bioluminescent labels are another area of nonisotopic immunoassays that continue to undergo active research. Most common approaches in this category are the competitive binding chemiluminescence immunoassays and the immunochemiluminometric assays. Chemiluminescence and heterogenous chemiluminescence immunoassays have been the subject of excellent reviews (91, 92). Detection in chemiluminescence immunoassays is based on either the direct monitoring of conjugated labels, such as luminol or acridinium ester, or the enzyme-mediated formation of luminescent products. Preparation of various derivatives of acridinium esters has been reported (93, 94), whereas a variety of enzyme labels including firefly or bacterial luciferase (70), horseradish peroxidase (86, 98), and alkaline phosphatase are commercially available. 

Bacterial luciferase High sensitivity. Needs no addition of substrate. No endogenous activity in mammalian cells. Heat sensitivity (>30°C). weak linearity... 

Lin L, Szitmer R, Meighen E. Binding of flavin and aldehyde to the active site of bacterial luciferase. In Stanley P, Kricka L. eds. Bioluminescence Chemiluminescence Progress Current Applications. Singapore World Scientific, 2002 89-92. 

In this work the interaction of bacterial luciferase from Photobacterium leiognathi with the flavin mononucleotide activated on phosphate group by N-methylimidazole, without addition of the exogenous aldehyde was study. 

Paralogues of eubacterial riboflavin synthase (designated as lumazine protein, yellow fluorescent protein, and blue fluorescent protein) without enzymatic activity have been isolated from several luminescent bacteria. These proteins are highly fluorescent and are believed to modulate to serve as optical transponders that modulate the spectral characteristics of bacterial bioluminescence.Energy is believed to be transferred from activated bacterial luciferase to the fluorescent proteins by radiation-less transfer. 

Bacterial luciferase is a heterodimer (75 kDa). The connection to the substrate is realized by the a-subunit (40 kDa), while the P-subunit (35 kDa) assures the stability of the active dimmer-structure. The luciferase molecule has active highly specific connection centers for FMN2, FMN, AMP and aldehyde [11]. 

It is well known that ATP is a major physiological parameter in cells, so that firefly luciferase is used for BL detection of bacterial contaminations, activity estimations of erythrocytes after blood conservation and antibiotic effects. BL measurements of microorganism growth rates is applied to assess antibiotic substrates, where the level of inhibition is most important. Information about the appropriate antibiotic treatment of open infections is received within 2-3 hours [28]. 

Heat The aj8 heterodimeric bacterial luciferases display 50% loss of activity within 8 min at45°C in 0.1 M phosphate buffer (pH 7.0) containing 1 mM EDTA and 0.1% BSA but not substrates (2). 

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