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Literature summary for 3.4.22.34 extracted from
- Comparative molecular analysis of two asparaginyl endopeptidases and encoding genes from Fasciola gigantica (2007), Mol. Biochem. Parasitol., 156, 102-116.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| inserted in pET20b(+), transformed into the Escherichia coli BL21(DE3)pLysS strain | Fasciola gigantica |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 1,10-phenanthroline | concentration of 1 mM, 10 min before addition of substrate, 40% inhibition; concentration of 1mM, 10 min before addition of substrate, 40% inhibition observed | Fasciola gigantica |  |
| benzyloxycarbonyl-L-Ala-L-Ala-azaAsn-chloromethylketone | Z-Ala-Ala-AzaAsn-CMK, 1 microM, 10 min before addition of substrate, 100% inhibition; Z-Ala-Ala-AzaAsn-CMK, 1 microM, 10 min before addition of substrate, 100% inhibition observed | Fasciola gigantica | |
| iodoacetamide | concentration of 5 mM,10 min before addition of substrate, 100% inhibition; concentration of 5 mM, 10 min before addition of substrate, 100% inhibition determined | Fasciola gigantica | |
| additional information | no significant inhibition by trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64) in a concentration of 10 microM, leupeptin in a concentration of 10 microM or pepstatin in a concentration of 5 microM; no significant inhibition observed by using trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64) in a concentration of 10 microM, leupeptin in a concentration of 10 microM or pepstatin in a concentration of 5 microM | Fasciola gigantica | |
| phenylmethanesulfonyl fluoride | PMSF, concentration of 2 mM, 10 min before addition of substrate, 66% inhibition; PMSF, concentration of 2 mM, 10 min before addition of substrate, 66% inhibition shown | Fasciola gigantica | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| intracellular | crude worm protein extracts but not in excretion-secretion products of adult parasites, polyclonal antisera and activity assay | Fasciola gigantica | 5622 | - |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Fasciola gigantica | A6Y9U8 | legumain-1, differentiated from legumain-2 by N-terminal sequence; collected from bile ducts and gall bladders of naturally infected cattle | - |
| Fasciola gigantica | A6Y9U9 | legumain-2, differentiated from legumain-1 by N-terminal sequence; collected from bile ducts and gall bladders of naturally infected cattle | - |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| proteolytic modification | N-terminal and C-terminal cleavage sites predicted, N-terminal sequence | Fasciola gigantica |
| proteolytic modification | N-terminal and C-terminal cleavage sites predicted, N-terminal sequence indicated | Fasciola gigantica |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli, gel filtration and SDS-PAGE | Fasciola gigantica |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| intestinal epithelium | caecal epithelium, 2-6 week old juveniles and adults, mRNA expression also in metacercariae | Fasciola gigantica | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
activity profile shown in crude worm protein extracts but not in excretion-secretion products of adult parasites, inhibition assay described, exon boundaries, landmark residues/motifs, and target areas for N- and C-terminal processing indicated | Fasciola gigantica |
| additional information | - |
activity profile shown in crude worm protein extracts but not in excretion-secretion products of adult parasites, inhibition assay, exon boundaries, landmark residues/motifs, and target areas for N- and C-terminal processing | Fasciola gigantica |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| benzoyl-Asn-4-nitroanilide + H2O | Bz-Asn-pNA, recombinant protein, at 37°C | Fasciola gigantica | benzoyl-Asn + 4-nitroaniline | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AEP | - |
Fasciola gigantica |
| Asparaginyl endopeptidase | - |
Fasciola gigantica |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
activity assay at | Fasciola gigantica |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5.4 | - |
activity assay at, maximum of activity observed at pH 5-6 | Fasciola gigantica |
| 5.4 | - |
activity assay at, maximum of activity observed at pH 56 | Fasciola gigantica |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 2 | 7 | assay under different pH conditions tested in 0.2 M sodium citrate/citric acid, 1 mM DTT at 37°C, no enzyme activity observed at pH 4 and below, activity dropped to 55% at pH 7 | Fasciola gigantica |
| 2 | 7 | assay under different pH conditions tested in 0.2 M sodium citrate/citric acid, 1 mM DTT at 37°C, no enzyme activity observed at pH 4 and below, activity drops to 55% at pH 7 | Fasciola gigantica |
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