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Literature summary for 1.13.12.13 extracted from
- Crystal structure of nanoKAZ The mutated 19kDa component of Oplophorus luciferase catalyzing the bioluminescent reaction with coelenterazine (2016), Biochem. Biophys. Res. Commun., 470, 88-93 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| recombinant expression of His-tagged 19 kDa catalytic subunit mutant nanoKAZ in Escherichia coli strain BL21. In bacterial cells, wild-type KAZ is mainly expressed as inclusion bodies | Oplophorus gracilirostris |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified mutated 19 kDa protein nanoKAZ, X-ray diffraction structure determination and analysis at 1.71-2.3.A resolution and dialysis | Oplophorus gracilirostris |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A4E/Q11R/Q18L/L27V/A33N/K43R/V44I/A54I/F68D/L72Q/M75K/I90V/P115E/Q124K/Y138I/N166R | construction of mutant nanoKAZ, a mutant with 16 amino acid substitutions of the catalytic subunit, by site-directed mutagenesis, the mutant nanoKAZ lacks the amino-terminal signal peptide, crystal structure determination and analysis, structure-function relationship in nanoKAZ, overview. The truncation of 10 amino acid residues at N- and C-terminal regions of nanoKAZ causes a complete loss of luminescence activity. Both the alpha1-helix and beta11-strand in the nanoKAZ molecule might serve to stabilize the molecule, which is essential in catalyzing the luminescence reaction | Oplophorus gracilirostris |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| extracellular | the enzyme possesses an amino-terminal signal peptide and is secreted | Oplophorus gracilirostris | - |
- |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 106000 | - |
full-length enzyme | Oplophorus gracilirostris |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| coelenterazine + O2 | Oplophorus gracilirostris | - |
coelenteramide + CO2 + hv | - |
? |  |
| Oplophorus luciferin + O2 | Oplophorus gracilirostris | - |
oxidized Oplophorus luciferin + CO2 + hv | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Oplophorus gracilirostris | Q9GV45 AND Q9GV46 | catalytic and noncatalytic subunit | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged catalytic subunit mutant nanoKAZ from Escherichia coli strain BL21 by nickel affinity chromatography and dialysis, wild-typeKAZ is purified from inclusion bodies using 6 M urea-nickel chelate affinity column chromatography | Oplophorus gracilirostris |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| wild-typeKAZ is purified from inclusion bodies using 6 M urea-nickel chelate affinity column chromatography | Oplophorus gracilirostris |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| coelenterazine + O2 | - |
Oplophorus gracilirostris | coelenteramide + CO2 + hv | - |
? | |
| coelenterazine + O2 | a luciferin substrate | Oplophorus gracilirostris | coelenteramide + CO2 + hv | - |
? | |
| Oplophorus luciferin + O2 | - |
Oplophorus gracilirostris | oxidized Oplophorus luciferin + CO2 + hv | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| heterooligomer | x * 19000, catalytic subunit, + x * 35000, noncatalytic subunit, SDS-PAGE | Oplophorus gracilirostris |
| More | the structure of the mutant catalytic subunit nanoKAZ consists of 11 antiparallel beta-strands forming a beta-barrel that is capped by 4 short alpha-helices. The coelenterazine-binding site and the catalytic site for the luminescence reaction might be in a central cavity of the beta-barrel structure of the catalytic subunit KAZ | Oplophorus gracilirostris |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| KAZ | catalytic enzyme subunit | Oplophorus gracilirostris |
| Oplophorus luciferase | - |
Oplophorus gracilirostris |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme is a member of the coelenterazine-utilizing luciferases such as Renilla luciferase and Gaussia luciferase | Oplophorus gracilirostris |
| additional information | the coelenterazine-binding site and the catalytic site for the luminescence reaction might be in a central cavity of the beta-barrel structure of the catalytic subunit KAZ | Oplophorus gracilirostris |
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