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Literature summary for 3.4.22.41 extracted from

  • Kang, J.M.; Bahk, Y.Y.; Cho, P.Y.; Hong, S.J.; Kim, T.S.; Sohn, W.M.; Na, B.K.
    A family of cathepsin F cysteine proteases of Clonorchis sinensis is the major secreted proteins that are expressed in the intestine of the parasite (2010), Mol. Biochem. Parasitol., 170, 7-16.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli M15 (pREP4) cells Clonorchis sinensis

Inhibitors

Inhibitors Comment Organism Structure
E-64 effective inhibition Clonorchis sinensis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
24000
-
fully matured enzymes CF-4 and CF-11, SDS-PAGE Clonorchis sinensis

Organism

Organism UniProt Comment Textmining
Clonorchis sinensis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography, gel filtration Clonorchis sinensis

Renatured (Commentary)

Renatured (Comment) Organism
for isozyme CF-4, the purified recombinant protein (10 mg) is slowly added to 1 l of 100 mM Tris-HCl (pH 8.0) containing 1 mM EDTA, 250 mM L-arginine, 5 mM reduced glutathione, and 1 mM oxidized glutathione and gently stirred at 4°C for 20 h. Isozyme CF-11 is refolded in 100 mM Tris-HCl (pH 8.0) containing 1 mM EDTA, 250 mM L-arginine, 10 mM reduced glutathione, and 1 mM oxidized glutathione with the same method Clonorchis sinensis

Source Tissue

Source Tissue Comment Organism Textmining
intestine CFs are synthesized in the epithelial cells lining the parasite intestine and secreted into the intestinal lumen Clonorchis sinensis
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Albumin + H2O substrate of isozyme CF-4 at pH 6.5 and CF-11 at pH 5.5 but not at neutral pH Clonorchis sinensis ?
-
?
benzyloxycarbonyl-L-leucine-L-arginine-7-amido-4-methylcoumarin + H2O high catalytic activity with isozymes CF-4 and CF-11 Clonorchis sinensis benzyloxycarbonyl-L-leucine-L-arginine + 7-amino-4-methylcoumarin
-
?
benzyloxycarbonyl-L-phenylalanine-L-arginine-7-amido-4-methylcoumarin + H2O high catalytic activity with isozymes CF-4 and CF-11 Clonorchis sinensis benzyloxycarbonyl-L-phenylalanine-L-arginine + 7-amino-4-methylcoumarin
-
?
Collagen + H2O substrate of isozyme CF-4 at pH 6.5 and CF-11 at pH 5.5 but not at neutral pH Clonorchis sinensis ?
-
?
Hemoglobin + H2O substrate of isozyme CF-4 at pH 6.5 and CF-11 at pH 5.5 but not at neutral pH Clonorchis sinensis ?
-
?
immunoglobulin A + H2O substrate of isozyme CF-4 at pH 6.5 and CF-11 at pH 5.5 but not at neutral pH Clonorchis sinensis ?
-
?
immunoglobulin G + H2O substrate of isozyme CF-4 at pH 6.5 and CF-11 at pH 5.5 but not at neutral pH Clonorchis sinensis ?
-
?
immunoglobulin M + H2O substrate of isozyme CF-4 at pH 6.5 and CF-11 at pH 5.5 but not at neutral pH Clonorchis sinensis ?
-
?
additional information the isozymes CF-4, CF-6, and CF-11 do not hydrolyze benzyloxycarbonyl-L-arginine-L-arginine-7-amido-4-methylcoumarin and benzyloxycarbonyl-L-arginine-7-amido-4-methylcoumarin Clonorchis sinensis ?
-
?

Synonyms

Synonyms Comment Organism
CF-11 isoform Clonorchis sinensis
CF-4 isoform Clonorchis sinensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
pH optimum of isozyme CF-11 Clonorchis sinensis
6.5
-
pH optimum of isozyme CF-4 Clonorchis sinensis

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 8 isozyme CF-4 is relatively stable at slight acidic and neutral pHs, but is comparatively unstable in acidic pHs, isozyme CF-11 is stable at acidic pHs, but is highly unstable in neutral and alkaline pH values Clonorchis sinensis

Expression

Organism Comment Expression
Clonorchis sinensis the isozymes CF-4, CF-6, and CF-11 are expressed throughout various developmental stages of the parasite and the transcripts increased gradually in accordance with the maturation of the parasite up

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.3
-
benzyloxycarbonyl-L-leucine-L-arginine-7-amido-4-methylcoumarin isozyme CF-11, in 100 mM sodium phosphate (pH 5.5), at 37°C Clonorchis sinensis
6
-
benzyloxycarbonyl-L-phenylalanine-L-arginine-7-amido-4-methylcoumarin isozyme CF-11, in 100 mM sodium phosphate (pH 5.5), at 37°C Clonorchis sinensis
7.6
-
benzyloxycarbonyl-L-leucine-L-arginine-7-amido-4-methylcoumarin isozyme CF-4, in 100 mM sodium phosphate (pH 6.5), at 37°C Clonorchis sinensis
8.2
-
benzyloxycarbonyl-L-phenylalanine-L-arginine-7-amido-4-methylcoumarin isozyme CF-4, in 100 mM sodium phosphate (pH 6.5), at 37°C Clonorchis sinensis