Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli M15 (pREP4) cells | Clonorchis sinensis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
E-64 | effective inhibition | Clonorchis sinensis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
24000 | - |
fully matured enzymes CF-4 and CF-11, SDS-PAGE | Clonorchis sinensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clonorchis sinensis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography, gel filtration | Clonorchis sinensis |
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 | Comment | Organism | Textmining |
---|---|---|---|
intestine | CFs are synthesized in the epithelial cells lining the parasite intestine and secreted into the intestinal lumen | Clonorchis sinensis | - |
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 | Comment | Organism |
---|---|---|
CF-11 | isoform | Clonorchis sinensis |
CF-4 | isoform | Clonorchis sinensis |
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 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 |
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 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 |