Oplophorus-luciferin 2-monooxygenase
| Oplophorus-luciferin 2-monooxygenase | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Identifiers | |||||||||
| EC no. | 1.13.12.13 | ||||||||
| Databases | |||||||||
| IntEnz | IntEnz view | ||||||||
| BRENDA | BRENDA entry | ||||||||
| ExPASy | NiceZyme view | ||||||||
| KEGG | KEGG entry | ||||||||
| MetaCyc | metabolic pathway | ||||||||
| PRIAM | profile | ||||||||
| PDB structures | RCSB PDB PDBe PDBsum | ||||||||
| |||||||||
Oplophorus-luciferin 2-monooxygenase (EC 1.13.12.13), also known as Oplophorus luciferase (referred in this article as OpLuc) is a luciferase, an enzyme, from the deep-sea shrimp Oplophorus gracilirostris,[1] belonging to a group of coelenterazine luciferases. Unlike other luciferases, it has a broader substrate specificity and can also bind to bisdeoxycoelenterazine efficiently.[1][2][3] It was the third example of a luciferase (Other than Aequorea and Renilla) to be purified in lab.[1] The systematic name of this enzyme class is Oplophorus-luciferin:oxygen 2-oxidoreductase (decarboxylating). This enzyme is also called Oplophorus luciferase.
Chemical reaction
The two substrates of this enzyme are the luciferin, coelenterazine and oxygen. Its products are the oxyluciferin, coelenteramide, carbon dioxide, and a photon. It belongs to the family of oxidoreductases, specifically those acting on single donors with O2 as oxidant and initial incorporation of two atoms of oxygen into the substrate. Although the enzyme is part of the group of enzymes that act on coelenterazine, such as Renilla and Gaussia luciferases, it does not share base pair sequences with these enzymes.[1][2][4][5]
OpLuc catalyzes the ATP independent chemical reaction:[6]
The result of this process is loss of CO2 and emission of a photon of blue light at ~460 nm.[1][2] This reaction has an optimal pH of 9, optimal salt concentration of 0.05-0.1 M, and optimal temperature of ~40 C (making it an unusually heat resistant luciferase),[1] although because O.gracilirostris are deep sea animals living in below 20 C temperatures, luciferase is normally expressed and folded at low temperatures.[3]
Biological function
When stimulated in Oplophorus gracilirostris, OpLuc is secreted from the base of legs and antennae of the deep-sea shrimp as a defense mechanism. This mechanism causes O.gracilirostris release a luminous, bright blue luciferase cloud.[1] There are many species of shrimp which display similar bioluminescence.[7]
Structure
OpLuc is a complex of two covalently bonded protein subunits: two molecules of 19 kDa and two molecules of 35 kDa components, making it a heterotetrameric molecule.[1] The proteins signal the enzyme for secretion in luminescence, catalyzed by the protein 19 kDa.[1][2][5] The luciferase has many cysteine residues that stabilize the enzyme in extracellular environments using disulfide bonds.[4]
19 kDa Protein
This catalytic component of OpLuc has 196 amino acids with one cysteine in the carboxyl terminus and is distinct from proteins found in other luciferases.[1][2] The protein is made up of two domains with repetitive sequencing of Ia-c and Ila-d in the peptide chain.[2] It is thought to be the protein to cause the bioluminescent reaction of O.gracilirostris, but functions ineffectively without its larger, subunit counterpart.[1][2] Although the crystal structure of OpLec has yet to be completely analyzed and mapped, 19 kDa experimentally expressed in mammalian cells (regarded as KAZ).[5] The protein was isolated and mutated to catalyze a bright and sustained luminescent reaction to create an engineered luciferase, NanoLuc (NLuc), and a coelenterazine analogue (furimazine) to be used as a cellular reporter.[4][8]
35 kDa Protein
The lesser known component of the OpLuc enzyme has 320 amino acids with 11 cysteine and 5 leucine molecules. The amino terminus of the protein was experimentally concluded to begin at 39 amino acids. It is thought to stabilize 19 kDa and is not thought to be affect by substrate specificity, however its exact function is not known.[1][2][5]
References
- ^ a b c d e f g h i j k l Shimomura, Osamu; Masugi, Takashi; Johnson, Frank H.; Haneda, Yata (1978). "Properties and reaction mechanism of the bioluminescence system of the deep-sea shrimp Oplophorus gracilorostris". Biochemistry. 17 (6): 994–998. doi:10.1021/bi00599a008. PMID 629957.
- ^ a b c d e f g h Inouye, Satoshi; Sasaki, Satoko (2007). "Overexpression, purification and characterization of the catalytic component of Oplophorus luciferase in the deep-sea shrimp, Oplophorus gracilirostris". Protein Expression and Purification. 56 (2): 261–268. doi:10.1016/j.pep.2007.08.002. PMID 17900925.
- ^ a b Inouye, Satoshi; Sahara-Miura, Yuiko; Sato, Jun-Ichi; Iimori, Rie; Yoshida, Suguru; Hosoya, Takamitsu (2013). "Expression, purification and luminescence properties of coelenterazine-utilizing luciferases from Renilla, Oplophorus and Gaussia: Comparison of substrate specificity for C2-modified coelenterazines". Protein Expression and Purification. 88 (1): 150–156. doi:10.1016/j.pep.2012.12.006. PMID 23274053.
- ^ a b c Hall, Mary P.; Unch, James; Binkowski, Brock F.; Valley, Michael P.; Butler, Braeden L.; Wood, Monika G.; Otto, Paul; Zimmerman, Kristopher; Vidugiris, Gediminas; Machleidt, Thomas; Robers, Matthew B.; Benink, Hélène A.; Eggers, Christopher T.; Slater, Michael R.; Meisenheimer, Poncho L.; Klaubert, Dieter H.; Fan, Frank; Encell, Lance P.; Wood, Keith V. (2012). "Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate". ACS Chemical Biology. 7 (11): 1848–1857. doi:10.1021/cb3002478. PMC 3501149. PMID 22894855.
- ^ a b c d Inouye, Satoshi; Sato, Jun-Ichi; Sahara-Miura, Yuiko; Hosoya, Takamitsu; Suzuki, Takahiro (2014). "Unconventional secretion of the mutated 19kDa protein of Oplophorus luciferase (NanoKAZ) in mammalian cells". Biochemical and Biophysical Research Communications. 450 (4): 1313–1319. doi:10.1016/j.bbrc.2014.06.140. PMID 25019994.
- ^ Enzyme 1.13.12.13 at KEGG Pathway Database.
- ^ Collins, Stormie B.; Bracken-Grissom, Heather D. (2024). "The language of light: A review of bioluminescence in deep-sea decapod shrimps". Biological Reviews. 99 (5): 1806–1830. doi:10.1111/brv.13093. PMID 38706106.
- ^ Tomabechi, Yuri; Hosoya, Takamitsu; Ehara, Haruhiko; Sekine, Shun-Ichi; Shirouzu, Mikako; Inouye, Satoshi (2016). "Crystal structure of nanoKAZ: The mutated 19 kDa component of Oplophorus luciferase catalyzing the bioluminescent reaction with coelenterazine". Biochemical and Biophysical Research Communications. 470 (1): 88–93. doi:10.1016/j.bbrc.2015.12.123. PMID 26746005.