Sandbox Reserved 1236
From Proteopedia
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== Function == | == Function == | ||
Luciferase | Luciferase | ||
| - | Luciferase is | + | Luciferase is a class of bioluminescent enzymes that are found in several luminescence organisms. The most studied form Luciferase within its class is found in the North American Firefly (Photinus pyralis). This protein catalyzed the reaction that produces the distinctive yellow flash seen in from the abdomen of the insect. Photinus pyralis is known to use this mechanism for mate attraction and defense. Firefly Luciferase is unique to its species and different forms within the luciferase class can be found in other invertebrates and bacteria.<ref>Conti, E., Franks, N. P., & Brick, P. (1996). Crystal structure of firefly luciferase throws light on a superfamily of adenylate-forming enzymes. Structure, 4(3), 287-298.</ref>. The most notable being the bioluminescent enzyme found in Click Beetles (Pyrophorus). This protein is one of the most studied and widely used luminescence enzymes having various applications in cell and molecular biology. |
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| + | =Origin== | ||
| + | Luciferase is part of the acyl-adenylate/thioester-forming superfamily. Through its two step reaction it ultimately forms an acyl-adenylate intermediate which is esterified into CoA. This mechanism is similar to the other members in this class some of which include Fatty Acyl AMP Ligases, Fatty Acyl CoA Ligases, and Acetyl CoA Ligases. There is evidence that Luciferase has homology with the highly conserved enzymes used in lipid synthesis. <ref>Khurana, Pankaj, Rajesh S. Gokhale, and Debasisa Mohanty. "Genome scale prediction of substrate specificity for acyl adenylate superfamily of enzymes based on active site residue profiles." BMC Bioinformatics 11.1 (2010): 57. ResearchGate. Web. 28 Mar. 2017.</ref> | ||
== Disease == | == Disease == | ||
== Relevance == | == Relevance == | ||
| - | This protein has been utilized in various types of assays to quantify the amount of ATP and to analyze the rate of transcription within a cell<ref>Wet, J. R., Wood, K. V., Helinski, D. R., & Deluca, M. (1985). Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli. Proceedings of the National Academy of Sciences, 82(23), 7870-7873. </ref>. This molecule is especially unique due to the fact that is very efficient in producing a photon through | + | This protein has been utilized in various types of assays to quantify the amount of ATP and to analyze the rate of transcription within a cell<ref>Wet, J. R., Wood, K. V., Helinski, D. R., & Deluca, M. (1985). Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli. Proceedings of the National Academy of Sciences, 82(23), 7870-7873. </ref>. This molecule is especially unique due to the fact that is very efficient in producing a photon through this reaction. It has potential for further biological applications in the future. Luciferase is widely used as a luminescent reporter gene in a variety of assays. The Luciferase gene can be isolated from the firefly and be inserted into animal or bacterial cells to monitor gene expression. This enzyme is sensitive to |
== Structural highlights == | == Structural highlights == | ||
The structure of this protein comprises of two prominent domains. The larger one contains an N terminal distorted beta-barrel accompanied by alpha helices. The second and smaller unit is consist of a beta sheet and alpha helix complex <ref>Viviani, V. R. (2002). The origin, diversity, and structure function relationships of insect luciferases. Cellular and Molecular Life Sciences, 59(11), 1833-1850. </ref>. The process of fluorescence is achieved through a two-step oxidation reaction involving the substrate Lucinferin accompanied with ATP, Magnesium and oxygen. The first step consist of using ATP-Mg in an Acylation reaction of the COOH group on Lucinferin producing a Luciferyl adenylate intermediate and a phosphate group. The second reaction uses oxygen to create an excited state of the molecule. The molecule then returns to its ground state emitting a photon of light (Conti et al., 1996). A single peptide has been discover that plays a vital role in the photooxidation by Luciferase. The specific amino acid is a histidine located in the region 244HHGF247 of the protein <ref>Branchini, B. R., Magyar, R. A., Marcantonio, K. M., Newberry, K. J., Stroh, J. G., Hinz, L. K., & Murtiashaw, M. H. (1997). Identification of a Firefly Luciferase Active Site Peptide Using a Benzophenone-based Photooxidation Reagent. Journal of Biological Chemistry, 272(31), 19359-19364.</ref>. It has been shown to be necessary for the use of oxygen in the second part of the reaction. | The structure of this protein comprises of two prominent domains. The larger one contains an N terminal distorted beta-barrel accompanied by alpha helices. The second and smaller unit is consist of a beta sheet and alpha helix complex <ref>Viviani, V. R. (2002). The origin, diversity, and structure function relationships of insect luciferases. Cellular and Molecular Life Sciences, 59(11), 1833-1850. </ref>. The process of fluorescence is achieved through a two-step oxidation reaction involving the substrate Lucinferin accompanied with ATP, Magnesium and oxygen. The first step consist of using ATP-Mg in an Acylation reaction of the COOH group on Lucinferin producing a Luciferyl adenylate intermediate and a phosphate group. The second reaction uses oxygen to create an excited state of the molecule. The molecule then returns to its ground state emitting a photon of light (Conti et al., 1996). A single peptide has been discover that plays a vital role in the photooxidation by Luciferase. The specific amino acid is a histidine located in the region 244HHGF247 of the protein <ref>Branchini, B. R., Magyar, R. A., Marcantonio, K. M., Newberry, K. J., Stroh, J. G., Hinz, L. K., & Murtiashaw, M. H. (1997). Identification of a Firefly Luciferase Active Site Peptide Using a Benzophenone-based Photooxidation Reagent. Journal of Biological Chemistry, 272(31), 19359-19364.</ref>. It has been shown to be necessary for the use of oxygen in the second part of the reaction. | ||
This is a sample scene created with SAT to <scene name="/12/3456/Sample/1">color</scene> by Group, and another to make <scene name="/12/3456/Sample/2">a transparent representation</scene> of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes. | This is a sample scene created with SAT to <scene name="/12/3456/Sample/1">color</scene> by Group, and another to make <scene name="/12/3456/Sample/2">a transparent representation</scene> of the protein. You can make your own scenes on SAT starting from scratch or loading and editing one of these sample scenes. | ||
| + | ==Applications== | ||
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</StructureSection> | </StructureSection> | ||
== References == | == References == | ||
<references/> | <references/> | ||
Revision as of 04:32, 29 March 2017
| This Sandbox is Reserved from Jan 17 through June 31, 2017 for use in the course Biochemistry II taught by Jason Telford at the Maryville University, St. Louis, USA. This reservation includes Sandbox Reserved 1225 through Sandbox Reserved 1244. |
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Luciferase
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References
- ↑ Conti, E., Franks, N. P., & Brick, P. (1996). Crystal structure of firefly luciferase throws light on a superfamily of adenylate-forming enzymes. Structure, 4(3), 287-298.
- ↑ Khurana, Pankaj, Rajesh S. Gokhale, and Debasisa Mohanty. "Genome scale prediction of substrate specificity for acyl adenylate superfamily of enzymes based on active site residue profiles." BMC Bioinformatics 11.1 (2010): 57. ResearchGate. Web. 28 Mar. 2017.
- ↑ Wet, J. R., Wood, K. V., Helinski, D. R., & Deluca, M. (1985). Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli. Proceedings of the National Academy of Sciences, 82(23), 7870-7873.
- ↑ Viviani, V. R. (2002). The origin, diversity, and structure function relationships of insect luciferases. Cellular and Molecular Life Sciences, 59(11), 1833-1850.
- ↑ Branchini, B. R., Magyar, R. A., Marcantonio, K. M., Newberry, K. J., Stroh, J. G., Hinz, L. K., & Murtiashaw, M. H. (1997). Identification of a Firefly Luciferase Active Site Peptide Using a Benzophenone-based Photooxidation Reagent. Journal of Biological Chemistry, 272(31), 19359-19364.
