Cloned (Comment) | Organism |
---|---|
expression and secretion of wild-type and mutants from Neuro2a cells | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
C108S | oligonucleotide-directed mutagenesis, reduced Km, increased kcat, increased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
C145S | oligonucleotide-directed mutagenesis of disulfide bond SS3, reduced Km and kcat, increased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
C208S | oligonucleotide-directed mutagenesis of disulfide bond SS6, highly reduced Km and reduced kcat, increased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
C233S | oligonucleotide-directed mutagenesis, highly reduced Km and slightly reduced kcat, reduced activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
C246S | oligonucleotide-directed mutagenesis, reduced Km and kcat, decreased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
C39S | oligonucleotide-directed mutagenesis of disulfide bond SS1, increased Km, reduced kcat, reduced activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
C7S | oligonucleotide-directed mutagenesis, reduced Km and kcat, slightly decreased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
D206V | oligonucleotide-directed mutagenesis, reduced Km and highly reduced kcat, decreased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-7-amino-4-methylcoumarin | Mus musculus |
additional information | additional deletion N113-E115 in mutant N110S: decreased kcat and Km, increased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
N110A | oligonucleotide-directed mutagenesis, reduced Km and kcat, slightly increased activity with substrate tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.22 | - |
tert-butyloxycarbonyl-Phe-Ser-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus | |
0.28 | - |
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus | |
0.32 | - |
tert-butyloxycarbonyl-Asp(benzyloxy)-Pro-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus | |
8.36 | - |
Pro-Phe-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | secreted | Mus musculus | - |
- |
extracellular | disruption of loop C and the kallikrein loop enhances the regulated secretion, no loop disruption does lead to inhibition of secretion | Mus musculus | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
polypeptide + H2O | Mus musculus | - |
peptides | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | Q61955 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | N-glycosylated at the kallikrein loop, residues His91-Ile103, oligosacchride chain is essential for activity | Mus musculus |
proteolytic modification | recombinant enzyme is produced as inactive proform and needs to be processed by an endoprotease, e.g. protease-1, EC 3.4.21.50, or trypsin, EC 3.4.21.4, for activation by specific cleavage of the Lys32-Ile33 bond near the N-terminus | Mus musculus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
brain | - |
Mus musculus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | recombinant enzyme is produced as inactive proform and needs to be processed by an endoprotease, e.g. protease-1, EC 3.4.21.50, or trypsin, EC 3.4.21.4, for activation by specific cleavage of the Lys32-Ile33 bond near the N-terminus | Mus musculus | ? | - |
? | |
additional information | loop C and the N-linked oligosaccharide chain on the kallikrein loop affect the catalytic efficiency and P2 specificity, respectively | Mus musculus | ? | - |
? | |
additional information | disulfide bonds SS1 in loop E, Gly142-Leu155, and SS6 in loop G, Ser185-Gly197, are essential for catalytic activity | Mus musculus | ? | - |
? | |
polypeptide + H2O | - |
Mus musculus | peptides | - |
? | |
Pro-Phe-Arg-4-methylcoumaryl-7-amide | recombinant wild-type and mutants | Mus musculus | Pro-Phe-Arg + 7-amino-4-methylcoumarin | - |
? | |
tert-butyloxycarbonyl-Asp(benzyloxy)-Pro-Arg-4-methylcoumaryl-7-amide + H2O | wild-type and mutants | Mus musculus | tert-butyloxycarbonyl-Asp(benzyloxy)-Pro-Arg + 7-amino-4-methylcoumarin | - |
? | |
tert-butyloxycarbonyl-Phe-Ser-Arg-4-methylcoumaryl-7-amide + H2O | recombinant wild-type and mutants | Mus musculus | tert-butyloxycarbonyl-Phe-Ser-Arg + 7-amino-4-methylcoumarin | - |
? | |
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide + H2O | best substrate | Mus musculus | tert-butyloxycarbonyl-Val-Pro-Arg + 7-amino-4-methylcoumarin | - |
? | |
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide + H2O | recombinant wild-type and mutants | Mus musculus | tert-butyloxycarbonyl-Val-Pro-Arg + 7-amino-4-methylcoumarin | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | determination of loop structure and organization | Mus musculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
31.3 | - |
tert-butyloxycarbonyl-Asp(benzyloxy)-Pro-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus | |
34.8 | - |
tert-butyloxycarbonyl-Phe-Ser-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus | |
100 | - |
tert-butyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus | |
193 | - |
Pro-Phe-Arg-4-methylcoumaryl-7-amide | recombinant wild-type enzyme, pH 8.0, 25°C | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Mus musculus |