Cloned (Comment) | Organism |
---|---|
expression of mutants as inactive pro-proteins in Pichia pastoris | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified wild-type enzyme and enzyme mutants in complex with inhibitor chagasin, hanging drop vapor diffusion method at 18°C, for crystal form I: mixing of 0.001 ml of 5 mg/ml chagasin solution with 0.002 ml of protein solution containing 3 mg/ml cathepsin B, and 0.002 ml of precipitant solution containing 0.2 M NH4H2PO4, 20% PEG 3350, pH 4.6, 2 weeks, for crystal form II: mixing of 0.001 mL of 5 mg/ml chagasin solution with 0.0015 ml of protein solution containing 3 mg/ml cathepsin B, and 0.001 ml of precipitant solution containing 0.14 M NH4F, 14% PEG 3350, pH 6.2, 1 week, X-ray diffraction structure determination and analysis at 1.8-2.7 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C29A | inactive mutant | Homo sapiens |
C29A/H110A/S115A | an inactive, non-glycosylated cathepsin B mutant | Homo sapiens |
H110A/S115A | a non-glycosylated cathepsin B mutant | Homo sapiens |
S115A | a non-glycosylated cathepsin B mutant | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
chagasin | an inhibitor from Trypanosoma cruzi, Three residues from chagasin, Leu65, Gly66, and Ala67, interact with cathepsin B residues Gly74, Gly73, and Asn72, binding mode and structure of wild-type enzyme and non-glycosylated S115A and H110A/S115A cathepsin B mutants, modeling, overview | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.055 | - |
carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin | pH 6.5, recombinant enzyme | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Homo sapiens | autocatalytic activation of the inactive proenzyme | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | glycosylation at Ser115 | Homo sapiens |
proteolytic modification | autocatalytic activation of the purified proenzyme in 20 mM sodium acetate, pH 5.0, 1mM EDTA, for 5 days | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
carbobenzoxy-Phe-Ala-Arg-7-amido-4-methylcoumarin + H2O | a fluorogenic substrate | Homo sapiens | carbobenzoxy-Phe-Ala-Arg + 7-amino-4-methylcoumarin | - |
? | |
carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin + H2O | a fluorogenic substrate | Homo sapiens | carbobenzoxy-Phe-Arg + 7-amino-4-methylcoumarin | - |
? | |
additional information | autocatalytic activation of the inactive proenzyme | Homo sapiens | ? | - |
? |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
autocatalytic assay at | Homo sapiens |
6.5 | - |
enzyme assay at, substrate carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin | Homo sapiens |