Photinus pyralis Luciferases, Luciferins

The bioluminescence systems of Phengodidae (railroad worms) and Elateroidae (click beetles) are basically identical to that of Lampyridae (fireflies), requiring firefly luciferin, ATP, Mg2+ and a luciferase for light emission. However, there seem to be some differences. Viviani and Bechara (1995) reported that the spectra of the luminescence reactions measured with the luciferases of Brazilian fireflies (6 species) shift from the yellow-green range to the red range with lowering of the pH of the medium, like in the case of the Photinus pyralis luciferase (see Section 1.1.5), whereas the spectra... 

Some new luminescent and fluorescent reporters (some of them even non-substrate proteins ) are very attractive because of their easy and fast detection, explaining their current frequent use. The bacterial luciferase isolated from the Vibrio fischeri lux operon contains luxAB encoding the functional subunits and luxCDE for the synthesis and recycling of the aldehyde substrate (Prosser, 1996). Firefly (Photinus pyralis) luciferase, encoded by the luc gene catalyses the oxidative carboxylation of beetle luciferin, in which photons are emitted (LaRossa, 1998). Its short half-life and lack of any post-translational modification makes it ideal to look after effects in gene expression (Naylor, 1999). Detection of... 

The most popular system in mechanistic and model studies as well as in analytical applications (clinical, food, environmental) appears to be that of firefly luciferin and luciferin-type-related model luminescence [3, 5,23, 57], The luciferase from Photinus pyralis, Photinus luciferin 4-monooxygenase (ATP-hydrolyzing), EC 1.13, 12.7, is a hydrophobic enzyme that catalyzes the air oxidation of luciferin in the presence of ATP and magnesium ions to yield light emission ... 

The reporter gene is the luc operon from the firefly Photinus pyralis. This operon codes for an enzyme, luciferase, that catalyses the reaction between D-luciferin (the substrate), and oxygen, and requires ATP as a cofactor. This reaction produces the emission light that provides the measure of enzyme activity. The oxidation of one molecule of luciferin involves one ATP molecule and releases one photon (a stoichiometric reaction) (DeLuca and McElroy, 1974 Wood el al. 1989). The reaction can be summarized as follows ... 

The firefly Photinus pyralis) is a familiar example of chemiluminescence. The photochemical reaction requires the enzyme luciferase (EC 1.13.12.7). In the presence of and the substrate luciferin, ATP is utilized in the following reaction... 

Although the total structure of the luciferase enzyme of the American firefly Photinus pyralis is unknown, the structure of the luciferin has been established. In... 

V. Bioluminescence-enhanced assay. This assay, originally developed for im-munoblot detection (20), utilizes coupled enzyme reactions initiated by APase that releases D-luciferin from D-luciferin O-phosphate (Novabiochem AG). The D-luciferin subsequently reacts with ATP and oxygen in a reaction catalyzed by luciferase (from firefly Photinus pyralis), resulting in light emission that can be detected on a sensitive photographic film (see Section D, Chapter 8). The reaction is performed in a pH 8.0 buffer. In immunoblot detections, this assay achieves a sensitivity of 5—50 pg protein (or 2—20 x 10 IgG molecules). 

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