Application | Comment | Organism |
---|---|---|
analysis | Cypridina noctiluca luciferase is utilized for biochemical and molecular biological applications, including bioluminescent enzyme immunoassays, far-red luminescence imaging, and high-throughput reporter assays | Cypridina noctiluca |
molecular biology | Cypridina noctiluca luciferase is utilized for biochemical and molecular biological applications, including bioluminescent enzyme immunoassays, far-red luminescence imaging, and high-throughput reporter assays | Cypridina noctiluca |
Cloned (Comment) | Organism |
---|---|
recombinant expression from vector p35SHSPG in Nicotiana benthamiana BY-2 cells, luciferase is efficiently secreted from cells with its native signal peptide, two potential N-glycosylation sites are modified with plant-type oligosaccharide chains. Recombinant expression of the enzyme in Arabidopsis thaliana roots in the cell wall of intact cells and the apoplast of plasmolyzed cells but not in the protoplasm. Cluc contains its own secretion signal peptide, which results in efficient secretion and folding of Cluc | Cypridina noctiluca |
Protein Variants | Comment | Organism |
---|---|---|
additional information | for large scale enzyme production, a simple production procedure is established using plant cell cultures. The plant cell culture tobacco BY-2 efficiently secretes luciferase, which is easily purified using a simple one-step ion-exchange chromatography method. The production yield is 20-30 mg of luciferase per liter of culture medium. The method offers a cost-effective production for Cypridina luciferase | Cypridina noctiluca |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Cypridina noctiluca | |
0.00028 | - |
Cypridina luciferin | recombinant enzyme, pH and temperature not specified in the publication | Cypridina noctiluca | |
0.00045 | - |
Vargula hilgendorfii luciferin | recombinant enzyme, recombinant substrate, pH and temperature not specified in the publication | Cypridina noctiluca | |
0.00052 | - |
Vargula hilgendorfii luciferin | recombinant enzyme, native substrate, pH and temperature not specified in the publication | Cypridina noctiluca |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | the enzyme Cluc contains a secretion signal peptide, which results in efficient secretion and folding of Cluc | Cypridina noctiluca | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
Cypridina luciferin + O2 | Cypridina noctiluca | - |
oxidized Cypridina luciferin + CO2 + hv | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cypridina noctiluca | Q75R40 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | glycosylation pattern analysis using LCMS/MS. In the recombinnat enzyme, two potential N-glycosylation sites are modified with plant-type oligosaccharide chains. Partial deglycosylation (trimming) of recombinant Cluc by beta-N-acetylhexosaminidase, HexNAc residues at the non-reducing termini of glycopeptides (Pronase E-digested) are completely eliminated at both glycosylation sites after beta-N-acetylhexosaminidase treatment (at Asn182 and Asn404). The glycan compositions after treatment displays trimannosyl cores (Hex3HexNAc2)+/-Fuc+/-Xyl. Specifically, because the trimannosyl core is fully occupied by Fuc and Xyl on Asn404, only a single signal corresponding to the trimannosyl core +Fuc +Xyl remains after treatment. This glycan-trimmed recombinant protein still exhibits nearly the same level of activity as the unmodified glycoprotein | Cypridina noctiluca |
Purification (Comment) | Organism |
---|---|
recombinant secreted enzyme from Nicotiana benthamiana BY-2 cell medium by ultrafiltration, buffer exchange gel filtration, anion exchange chromatography, and dialysis | Cypridina noctiluca |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Cypridina luciferin + O2 | - |
Cypridina noctiluca | oxidized Cypridina luciferin + CO2 + hv | - |
? | |
Vargula hilgendorfii luciferin + O2 | native or recombinant substrate protein | Cypridina noctiluca | oxidized Vargula hilgendorfii luciferin + CO2 + hv | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 61000, recombinnat glycolysated enzyme, SDS-PAGE | Cypridina noctiluca |
Synonyms | Comment | Organism |
---|---|---|
CLuc | - |
Cypridina noctiluca |
Cypridina luciferase | - |
Cypridina noctiluca |