User:Matt Whelihan: Difference between revisions
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Luciferases have become an invaluable tool in microbiology and biochemistry as a means of reporting gene expression and in-vivo/vitro chemical conditions. Since it was first cloned in 1985 by DeLuca et al., the gene coding for luciferases has been used in reporter assays to measure gene transcription and cellular morphology. Various cell lines have also been engineered to express luciferases as a measure of the oxidative state in various organs and types of diseases. Perhaps its most useful function is in the efficient detection of intracellular ATP. Luciferases are also used to detect protein-anesthetic interactions as they are susceptible to a wide range of general anesthetics. | Luciferases have become an invaluable tool in microbiology and biochemistry as a means of reporting gene expression and in-vivo/vitro chemical conditions. Since it was first cloned in 1985 by DeLuca et al., the gene coding for luciferases has been used in reporter assays to measure gene transcription and cellular morphology. Various cell lines have also been engineered to express luciferases as a measure of the oxidative state in various organs and types of diseases. Perhaps its most useful function is in the efficient detection of intracellular ATP. Luciferases are also used to detect protein-anesthetic interactions as they are susceptible to a wide range of general anesthetics. | ||
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== Firefly Luciferase == | == Firefly Luciferase == | ||
One of the most studied of all luciferases is that of the <scene name='User:Matt_Whelihan/Rainbow_n-c/2'>Firefly</scene>. This particular luciferase enzyme is located in the light emitting organ known as the lantern in the abdomen of the beetle. Firefly larvae glow green to ward off predators and adult fireflies use this mechanism of bioluminescence to attract mates. Luciferase binds ATP/Mg+ and D-luciferin and oxidizes it to Oxyluciferin with the products of one photon of yellow-green light, pyrophosphate, AMP and CO2<ref>PMID:775940</ref>. The crystal structures of two firefly luciferases, the North American Firefly (''Photinus pyralis'') <ref>PMID:8805533</ref> and the Japanese Firefly (''Luciola cruciata'')<ref>PMID:16541080</ref> 16541080 have been solved. | One of the most studied of all luciferases is that of the <scene name='User:Matt_Whelihan/Rainbow_n-c/2'>Firefly</scene>. This particular luciferase enzyme is located in the light emitting organ known as the lantern in the abdomen of the beetle. Firefly larvae glow green to ward off predators and adult fireflies use this mechanism of bioluminescence to attract mates. Luciferase binds ATP/Mg+ and D-luciferin and oxidizes it to Oxyluciferin with the products of one photon of yellow-green light, pyrophosphate, AMP and CO2<ref>PMID:775940</ref>. The crystal structures of two firefly luciferases, the North American Firefly (''Photinus pyralis'') <ref>PMID:8805533</ref> and the Japanese Firefly (''Luciola cruciata'')<ref>PMID:16541080</ref> 16541080 have been solved. | ||
<applet load='2d1r' size='400' frame='true' align='left' caption='Crystal structure of the thermostable Japanese firefly Luciferase complexed with OXYLUCIFERIN [http://www.rcsb.org/pdb/ligand/ligandsummary.do?hetId=OLU&sid=2D1R] and AMP [http://www.rcsb.org/pdb/ligand/ligandsummary.do?hetId=AMP&sid=2D1R]' /> | |||
Both the Japanese and North American luciferase variants consist of a single-chain 62 kDa monooxygenases with two distinct domains that are <scene name='User:Matt_Whelihan/Secondary_structure/1'>comprised</scene> of a mix of <font color='magenta'>alpha</font> and <font color='gold'>beta</font> secondary structures. The <scene name='User:Matt_Whelihan/N-terminal_domain/1'>N-terminal domain</scene> (residues 4-436) is comprised of of an antiparalell β-barrel and two β-sheets, flanked by α-helices which forms a typical <scene name='User:Matt_Whelihan/Rossmann/1'>Rossmann fold</scene> [http://images.google.com/imgres?imgurl=http://chem.csusb.edu/~dpedersn/C436/openbetarossman.jpg&imgrefurl=http://chem.csusb.edu/~dpedersn/C436/prot_struct_and_class.html&usg=__xOF4ifmAyJ5qm3UPaNKd9whKlu4=&h=539&w=719&sz=35&hl=en&start=5&um=1&tbnid=IMkWKmy0xzWcYM:&tbnh=105&tbnw=140&prev=/images%3Fq%3DRossman%2BFold%26hl%3Den%26safe%3Doff%26sa%3DN%26um%3D1] The | Both the Japanese and North American luciferase variants consist of a single-chain 62 kDa monooxygenases with two distinct domains that are <scene name='User:Matt_Whelihan/Secondary_structure/1'>comprised</scene> of a mix of <font color='magenta'>alpha</font> and <font color='gold'>beta</font> secondary structures. The <scene name='User:Matt_Whelihan/N-terminal_domain/1'>N-terminal domain</scene> (residues 4-436) is comprised of of an antiparalell β-barrel and two β-sheets, flanked by α-helices which forms a typical <scene name='User:Matt_Whelihan/Rossmann/1'>Rossmann fold</scene> [http://images.google.com/imgres?imgurl=http://chem.csusb.edu/~dpedersn/C436/openbetarossman.jpg&imgrefurl=http://chem.csusb.edu/~dpedersn/C436/prot_struct_and_class.html&usg=__xOF4ifmAyJ5qm3UPaNKd9whKlu4=&h=539&w=719&sz=35&hl=en&start=5&um=1&tbnid=IMkWKmy0xzWcYM:&tbnh=105&tbnw=140&prev=/images%3Fq%3DRossman%2BFold%26hl%3Den%26safe%3Doff%26sa%3DN%26um%3D1] The | ||
