(Redirected from Firefly luciferase)
'Luciferase' is a generic name for
enzymes commonly used in nature for
bioluminescence. The name itself is derived from ''
Lucifer'', which means ''light-bearer''. The most famous one is
firefly luciferase () from the firefly ''Photinus pyralis''. In luminescent reactions, light is produced by the
oxidation of a
luciferin (a pigment), sometimes involving
Adenosine triphosphate (ATP). The rates of this reaction between luciferin and
oxygen are extremely slow until they are
catalyzed by luciferase, often mediated by the presence of
calcium ions (an analog of
muscle contraction)
[1]. The reaction takes place in two steps:
luciferin + ATP → luciferyl adenylate + PPi
luciferyl adenylate + O2 → oxyluciferin + AMP + light
The reaction is very energy efficient: nearly all of the energy input into the reaction is transformed into light. As a comparison, the
incandescent light bulb loses about 90% of its
energy to
heat.
'Luciferin' and 'luciferase' are not specific molecules. They are generic terms for a substrate and its associated
enzyme (or
protein) that
catalyze a light-producing reaction. A variety of organisms regulate their light production using different luciferases in a variety of light-emitting reactions. The most famous are the
fireflies, although the enzyme exists in organisms as different as the Jack-O-Lantern mushroom ''(
Omphalotus olearius)'' and many marine creatures. In fireflies, the oxygen required is supplied through a tube in the abdomen called the
abdominal trachea. Some organisms, notably certain
click beetles, have several different luciferase enzymes, which each can produce different colors from the same luciferin. The luciferases of fireflies - of which there are over 2000
species - and of the
Elateroidea (fireflies, click beetles and relatives) in general - are diverse enough to be useful in
molecular phylogeny. The most thoroughly studied luciferase is that of the
Photinini firefly ''
Photinus pyralis''.
Applications
Luciferase can be produced in the lab through
genetic engineering for a number of purposes. Luciferase
genes can be synthesized and inserted into organisms or transfected into cells.
Mice,
silkworms, and
potatoes are just a few organisms that have already been engineered to produce the protein.
Ex-vivo imaging is a very powerful technique for studying cell populations in whole animals. Different types of cells (e.g. bone marrow stem cells, T-cells) can be engineered to express a luciferase allowing their non-invasive visualization inside a live animal using a sensitive
CCD camera.
Light is emitted when luciferase is exposed to the appropriate luciferin
substrate. Photon emission can be detected by light sensitive apparatus such as a
luminometer or modified optical microscopes. This allows observation of biological processes and stages of infection, for example. Luciferase can be used in
blood banks to determine if
red blood cells are starting to break down.
Forensic investigators can use a dilute solution containing the enzyme to uncover traces of blood remaining on surfaces at a crime scene. Laboratories can employ luciferase to emit light in the presence of certain
diseases. Luciferase is used as a reporter protein in molecular studies, for example to test the activity of
transcription from specific
promoters in cells transfected with luciferase, or to detect the level of cellular
ATP. Luciferase is a very heat sensitive protein that is used in studies on
protein denaturation, testing the protective capacities of
heat shock proteins. The opportunities for using luciferase continue to expand.
See also
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Firefly
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Quorum sensing
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Bioluminescence
References
1. IUBMB Enzyme Nomenclature ''Renilla'' luciferin reaction is triggered by calcium ions. (cited September 6, 2006)
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Thomas O Baldwin. Firefly luciferase: the structure is known, but the mystery remains Structure. 1996 Mar 15;4(3):223-8.
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Greer LF 3rd, Szalay AA. Imaging of light emission from the expression of luciferases in living cells and organisms: a review Luminescence. 2002 Jan-Feb;17(1):43-74.
External links
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Chemistry of Bioluminescence
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PDB for Firefly Luciferase
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PDB for Bacterial Luciferase
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Nitric oxide and firefly flashing
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Promega Gene Expression and Reporter technologies using Luciferase