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Literature summary for 3.4.22.B49 extracted from
- Cloning and characterization of a basic cysteine-like protease (cathepsin L1) expressed in the gut of larval Diaprepes abbreviatus L. (Coleoptera Curculionidae) (2015), J. Insect Physiol., 72, 1-13 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| cloned from digestive tracts of larva and adult in different stages, DNA and amino acid sequence determination and analysis of proCTSL1, recombinant expression of N-terminally GST-tagged and C-terminally His6-tagged enzyme in Escherichia coli strain Rosetta (DE3) | Diaprepes abbreviatus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Aprotinin | - |
Diaprepes abbreviatus |  |
| CA074 | 25.3% inhibition at 0.05 mM | Diaprepes abbreviatus | |
| chymostatin | 85.6% inhibition at 0.05 mM | Diaprepes abbreviatus | |
| cystatin | 47.5% inhibition at 0.05 mM | Diaprepes abbreviatus | |
| E-64 | 26.6% inhibition at 0.05 mM | Diaprepes abbreviatus | |
| N-benzyloxycarbonyl-FY(tert-butyl)-diazomethylketone | 40.6% inhibition at 0.05 mM | Diaprepes abbreviatus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Diaprepes abbreviatus | Q0PZI4 | i.e. Curculio abbreviatus | - |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| proteolytic modification | cleavage of the propeptide from proCTSL1 for enzyme maturation and activtaion | Diaprepes abbreviatus |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant GST- and His6-tagged proCTSL1 enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and gel filtration | Diaprepes abbreviatus |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| gut | - |
Diaprepes abbreviatus | - |
| larva | - |
Diaprepes abbreviatus | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | substrate specificity supports the hypothesis that Diaprepes abbreviatus CTSL1 is a unique basic cathepsin L and protease inhibitor studies also suggest unique activity, unlike other characterized acidic cathepsin Ls. No activity with N-succinyl-AAPF-4-nitroanilide and HR-4-nitroanilide | Diaprepes abbreviatus | ? | - |
? | |
| N-benzyloxycarbonyl-FR-4-nitroanilide + H2O | - |
Diaprepes abbreviatus | N-benzyloxycarbonyl-FR + 4-nitroaniline | - |
? | |
| N-benzyloxycarbonyl-FR-7-amido-4-trifluoromethylcoumarin + H2O | best substrate | Diaprepes abbreviatus | N-benzyloxycarbonyl-FR + 7-amino-4-trifluoromethylcoumarin | - |
? | |
| N-benzyloxycarbonyl-GPR-7-amido-4-methylcoumarin + H2O | - |
Diaprepes abbreviatus | N-benzyloxycarbonyl-GPR + 7-amino-4-methylcoumarin | - |
? | |
| N-benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin + H2O | - |
Diaprepes abbreviatus | N-benzyloxycarbonyl-VVR + 7-amino-4-methylcoumarin | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 60000, recombinant of N-terminally GST-tagged and C-terminally His6-tagged pro-enzyme, SDS-PAGE, x * 23000, recombinant detagged mature enzyme, SDS-PAGE | Diaprepes abbreviatus |
| More | three dimensional structure modeling of the protein sequence shows that the mature Da-CTSL1 protein folds into an expected cathepsin L structure producing a substrate binding pocket with appropriate positioning of conserved amino acid residues | Diaprepes abbreviatus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Da-CTSL1 | - |
Diaprepes abbreviatus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Diaprepes abbreviatus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
recombinant mature enzyme | Diaprepes abbreviatus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 5.5 | 10 | activity range, profile overview. 8Fold higher activity at pH 8.0 as compared to pH 5.5 with substrate N-benzyloxycarbonyl-GPR-7-amido-4-methylcoumarin, Da-CTSL1 is a mainly alkaline cathepsin | Diaprepes abbreviatus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | three-dimensional structure modeling of the protein sequence shows that the mature Da-CTSL1 protein folds into an expected cathepsin L structure producing a substrate binding pocket with appropriate positioning of conserved amino acid residues. The C-terminal catalytic domain possesses the two catalytically active Cys120 and His255 amino acid residues located in a pocket at the center of the predicted mature protein catalytic groove. Both domains are organized in such a way that the propeptide (PP) becomes inserted into the groove and completely masks the catalytic pocket of the active domain, inhibiting the proteinase activity of Da-CTSL1. The three-dimensional model also predicts that a network of fourteen hydrogen bonds involving D231, T233, L253, N254, E255, and E300 residues, allows the PP to anchor at the Da-CTSL1 catalytic groove. Residues R71, L76, S77, S78, S79, and K80 of PP participate in this H-bonding network | Diaprepes abbreviatus |
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