Cloned (Comment) | Organism |
---|---|
pseudozymogen forms of recombinant matriptase are expressed in Pichia pastoris and in CHO cells | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | engineered L-Matriptase is a secreted variant of r-matriptase in which the cytosolic, signal anchor, and stem domains (Met1-Asp603) are replaced with the human immunoglobulin kappa-chain signal peptide and S-tag (ST). L-Matriptase is produced in CHO-K1 cells and is converted to an active form via cleavage with recombinant enterokinase | Rattus norvegicus |
additional information | four matriptase pseudozymogens containing at least the catalytic domain are constructed. Each plasmid is expressed as His-tagged fusion proteins and contains a site for activation by recombinant enterokinase. Pseudozymogens with His6-tag at their C-termini form multimers linked by intermolecular disulfide bonds. After treatment with r-EK, they exhibit no detectable hydrolytic activity. Designated pseudozymogen His6t-S-CD consisting of a spacer and the catalytical domain with an N-terminal His6-tag is secreted as a monomer. After recombinant enterokinase treatment pseudozymogen His6t-S-CD exhibits enzymatic activity comparable to secreted variant L-matriptase | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide | pseudozymogen His6t-S-CD consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase is used | Rattus norvegicus | |
0.19 | - |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide | engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase) | Rattus norvegicus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
SDS-PAGE reducing conditions and Western blot using anti-rat matriptase catalytic domain anibody, designated pseudozymogen His6t-S-CD consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase, after cleavage with enterokinase | Rattus norvegicus |
32000 | - |
SDS-PAGE reducing and non-reducing conditions and Western blot using anti-rat matriptase catalytic domain anibody, designated pseudozymogen His6t-S-CD consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase, before cleavage with enterokinase | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
proteolytic modification | - |
Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-Lys-Thr-Lys-Gln-Leu-Arg-methyl-coumaryl-7-amide + H2O | relative activity: 19% using engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase), relative activity: 16% using engineered pseudozymogen His6t-S-CD (consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase) | Rattus norvegicus | acetyl-Lys-Thr-Lys-Gln-Leu-Arg + 7-amino-4-methylcoumarin | - |
? | |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide + H2O | - |
Rattus norvegicus | methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine + p-nitroaniline | - |
? | |
t-butyloxycarbonyl-Asp-Pro-Arg-methyl-coumaryl-7-amide + H2O | relative activity: 66% using engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase), relative activity: 69% using engineered pseudozymogen His6t-S-CD (consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase) | Rattus norvegicus | t-butyloxycarbonyl-Asp-Pro-Arg + 7-amino-4-methylcoumarin | - |
? | |
t-butyloxycarbonyl-Gln-Ala-Arg-methyl-coumaryl-7-amide + H2O | relative activity: 100%, using engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase) or engineered pseudozymogen His6t-S-CD (consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase) | Rattus norvegicus | t-butyloxycarbonyl-Gln-Ala-Arg + 7-amino-4-methylcoumarin | - |
? | |
t-butyloxycarbonyl-Glu-Gly-Arg-methyl-coumaryl-7-amide + H2O | relative activity: 29% using engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase), relative activity: 31% using engineered pseudozymogen His6t-S-CD (consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase) | Rattus norvegicus | t-butyloxycarbonyl-Glu-Gly-Arg + 7-amino-4-methylcoumarin | - |
? | |
t-butyloxycarbonyl-Leu-Gly-Arg-methyl-coumaryl-7-amide + H2O | relative activity: 28% using engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase), relative activity: 28% using engineered pseudozymogen His6t-S-CD (consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase) | Rattus norvegicus | t-butyloxycarbonyl-Leu-Gly-Arg + 7-amino-4-methylcoumarin | - |
? | |
t-butyloxycarbonyl-Phe-Ser-Arg-methyl-coumaryl-7-amide + H2O | relative activity: 12% using engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase), relative activity: 13% using engineered pseudozymogen His6t-S-CD (consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase) | Rattus norvegicus | t-butyloxycarbonyl-Phe-Ser-Arg + 7-amino-4-methylcoumarin | - |
? |
Synonyms | Comment | Organism |
---|---|---|
matriptase | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
59 | - |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide | engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase) | Rattus norvegicus | |
59 | - |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide | pseudozymogen His6t-S-CD consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase is used | Rattus norvegicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
310 | - |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide | engineered L-matriptase (secreted variant of matriptase activated by recombinant enterokinase) | Rattus norvegicus | |
350 | - |
methylsulfonyl-D-cyclohexyltyrosyl-glycyl-arginine-p-nitroanilide | pseudozymogen His6t-S-CD consisting of a spacer and the catalytical domain with an N-terminal His6-tag and a cleavage site for activation by enterokinase is used | Rattus norvegicus |