Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.21.109 extracted from

  • Mochida, S.; Tsuzuki, S.; Yasumoto, M.; Inouye, K.; Fushiki, T.
    Secreted expression of pseudozymogen forms of recombinant matriptase in Pichia pastoris (2009), Enzyme Microb. Technol., 45, 288-294.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
pseudozymogen forms of recombinant matriptase are expressed in Pichia pastoris and in CHO cells Rattus norvegicus

Protein Variants

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 [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 [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

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification
-
Rattus norvegicus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
matriptase
-
Rattus norvegicus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Rattus norvegicus

Turnover Number [1/s]

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 [mM/s]

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