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Literature summary extracted from

  • Inouye, S.; Sato, J.; Sahara-Miura, Y.; Yoshida, S.; Hosoya, T.
    Luminescence enhancement of the catalytic 19 kDa protein (KAZ) of Oplophorus luciferase by three amino acid substitutions (2014), Biochem. Biophys. Res. Commun., 445, 157-162 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.12.13 recombinant expression and secretion, using the signal peptide sequence of Gaussia luciferase, of catalytic 19 kDa protein KAZ of Oplophorus luciferase and its mutants in Escherichia coli and recombinant expression of His-tagged wild-type and mutant enzymes and KAZs in CHO-K1 cells Oplophorus gracilirostris

Protein Variants

EC Number Protein Variants Comment Organism
1.13.12.13 A33N site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 A4E site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows wild-type bioluminesce intensity Oplophorus gracilirostris
1.13.12.13 A54I site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 A54I/Y138I site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 F68D site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows wild-type bioluminesce intensity Oplophorus gracilirostris
1.13.12.13 I90V site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 K43R site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows wild-type bioluminesce intensity Oplophorus gracilirostris
1.13.12.13 L27V site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows reduced bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 L72Q site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 M75K site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 additional information construction of mutant nanoKAZ, a mutant with 16 amino acid substitutions of the catalytic subunit. Secretory expression and luminescence activity of single amino acid substituted KAZ mutants in CHO-K1 cells, overview Oplophorus gracilirostris
1.13.12.13 N166R site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 P115E site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 Q11R site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 Q124K site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 Q18L site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows wild-type bioluminesce intensity Oplophorus gracilirostris
1.13.12.13 V44I site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 V44I/A54I site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris
1.13.12.13 V44I/A54I/Y138I site-directed mutagenesis, mutation of the catalytic subunit, catalytic subunit mutant, that shows 7fold higher activity than 16-aa-mutant nanoKAZ using coelenterazine, but these substitutions does not stimulate protein secretion from mammalian cells, nanoKAZ possessing the signal peptide sequence of Gaussia luciferase for secretion expressed efficiently into the culture medium of CHO-K1 cells Oplophorus gracilirostris
1.13.12.13 V44I/Y138I site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows wild-type bioluminesce intensity Oplophorus gracilirostris
1.13.12.13 Y138I site-directed mutagenesis, mutation of the catalytic subunit, the mutant shows increased bioluminesce intensity compared to the wild-type Oplophorus gracilirostris

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.13.12.13 extracellular the enzyme is secreted Oplophorus gracilirostris
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.13.12.13 106000
-
 full-length enzyme Oplophorus gracilirostris

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.13.12.13 coelenterazine + O2 Oplophorus gracilirostris
-
 coelenteramide + CO2 + hv
-
 ?
1.13.12.13 additional information Oplophorus gracilirostris Oplophorus luciferase shows broad substrate specificities for various coelenterazine analogues, and the substrate specificity is distinct from other coelenterazine-type luciferases including Renilla and Gaussia luciferases and the Ca2+-binding photoprotein aequorin ?
-
 ?
1.13.12.13 Oplophorus luciferin + O2 Oplophorus gracilirostris
-
 oxidized Oplophorus luciferin + CO2 + hv
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.13.12.13 Oplophorus gracilirostris Q9GV45 AND Q9GV46 catalytic and noncatalytic subunit
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.13.12.13 recombinant His-tagged catalytic wild-type and mutant enzymes and catalytic subunits from Escherichia coli by nickel affinity chromatgraphy Oplophorus gracilirostris

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.12.13 6h-coelenterazine + O2
-
 Oplophorus gracilirostris ? + CO2 + hv
-
 ?
1.13.12.13 6h-f-coelenterazine + O2
-
 Oplophorus gracilirostris ? + CO2 + hv
-
 ?
1.13.12.13 bis-coelenterazine + O2
-
 Oplophorus gracilirostris ? + CO2 + hv
-
 ?
1.13.12.13 coelenterazine + O2
-
 Oplophorus gracilirostris coelenteramide + CO2 + hv
-
 ?
1.13.12.13 f-coelenterazine + O2
-
 Oplophorus gracilirostris ? + CO2 + hv
-
 ?
1.13.12.13 furimazine + O2
-
 Oplophorus gracilirostris ? + CO2 + hv
-
 ?
1.13.12.13 h-coelenterazine + O2
-
 Oplophorus gracilirostris ? + CO2 + hv
-
 ?
1.13.12.13 additional information Oplophorus luciferase shows broad substrate specificities for various coelenterazine analogues, and the substrate specificity is distinct from other coelenterazine-type luciferases including Renilla and Gaussia luciferases and the Ca2+-binding photoprotein aequorin Oplophorus gracilirostris ?
-
 ?
1.13.12.13 Oplophorus luciferin + O2
-
 Oplophorus gracilirostris oxidized Oplophorus luciferin + CO2 + hv
-
 ?

Subunits

EC Number Subunits Comment Organism
1.13.12.13 heterooligomer x * 19000, catalytic subunit, + x * 35000, noncatalytic subunit, SDS-PAGE Oplophorus gracilirostris

Synonyms

EC Number Synonyms Comment Organism
1.13.12.13 KAZ catalytic enzyme subunit Oplophorus gracilirostris
1.13.12.13 Oplophorus luciferase
-
 Oplophorus gracilirostris

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.13.12.13 7.6
-
 assay at Oplophorus gracilirostris

General Information

EC Number General Information Comment Organism
1.13.12.13 physiological function the secreted luciferase of the deep-sea shrimp Oplophorus gracilirostris catalyzes the oxidation of coelenterazine to emit blue light (lambda_max = 460 nm) Oplophorus gracilirostris