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Literature summary extracted from

  • Abdul Alim, M.; Tsuji, N.; Miyoshi, T.; Khyrul Islam, M.; Huang, X.; Motobu, M.; Fujisaki, K.
    Characterization of asparaginyl endopeptidase, legumain induced by blood feeding in the ixodid tick Haemaphysalis longicornis (2007), Insect Biochem. Mol. Biol., 37, 911-922.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.22.34 plasmid pTrcHisB/HlLgm transformed into the Escherichia coli Top10F0 strain Haemaphysalis longicornis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.34 egg white cystatin concentration of 2 microM, 45.13% inhibition of recombinant protein activity Haemaphysalis longicornis
3.4.22.34 iodoacetamide concentration of 3 mM, 100% inhibition of recombinant protein activity, BSA cleavage inhibited by 5 mM iodoacetamide Haemaphysalis longicornis
3.4.22.34 leupeptin concentration of 300 microM, 16.07% inhibition of recombinant protein activity Haemaphysalis longicornis
3.4.22.34 N-ethylmaleimide concentration of 3 mM, 95.74% inhibition of recombinant protein activity Haemaphysalis longicornis
3.4.22.34 pepstatin concentration of 500 microM, 7.8% inhibition of recombinant protein activity Haemaphysalis longicornis
3.4.22.34 phenylmethanesulfonyl fluoride PMSF, concentration of 5 mM, 19% inhibition of recombinant protein activity Haemaphysalis longicornis
3.4.22.34 trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane E-64, concentration of 500 microM, 8.88% inhibition of recombinant protein activity Haemaphysalis longicornis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.22.34 membrane bound Haemaphysalis longicornis 16020
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.22.34 38000
-
 removal of a peptide of 9000 Dalton from the preprotein Haemaphysalis longicornis
3.4.22.34 47000
-
 preprotein, deduced from sequence Haemaphysalis longicornis

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.34 Haemaphysalis longicornis A4PF00 encoded by the HlLgm gene; parthenogenetic Okayama strain
-
3.4.22.34 Haemaphysalis longicornis Okayama A4PF00 encoded by the HlLgm gene; parthenogenetic Okayama strain
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
3.4.22.34 proteolytic modification cleavage site at asparagines 364-365 at the C-terminus identified in endogenous and in recombinant protein Haemaphysalis longicornis

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.22.34 SDS-PAGE Haemaphysalis longicornis

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.4.22.34 midgut lmidgut epithelium shown by immunohistochemistry Haemaphysalis longicornis
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.22.34 0.642
-
 recombinant protein, 37°C, pH 7, benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin-7-amide, activity profile shown, inhibition assay described, up-regulation by the host blood-feeding process shown by in vitro proteolysis assay Haemaphysalis longicornis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.34 benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin-7-amide + H2O recombinant protein Haemaphysalis longicornis benzyloxycarbonyl-Ala-Ala-Asn + 7-amino-4-methylcoumarin
-
 ?
3.4.22.34 benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin-7-amide + H2O recombinant protein Haemaphysalis longicornis Okayama benzyloxycarbonyl-Ala-Ala-Asn + 7-amino-4-methylcoumarin
-
 ?
3.4.22.34 Bovine serum albumin + H2O degradation in a dose-dependent manner at pH 7 and 30°C during 6 h of incubation, no cleavage detected at or above 37°C, strict specificity for hydrolysis of the peptide on the carboxyl side of the asparagines shown Haemaphysalis longicornis ?
-
 ?
3.4.22.34 Bovine serum albumin + H2O degradation in a dose-dependent manner at pH 7 and 30°C during 6 h of incubation, no cleavage detected at or above 37°C, strict specificity for hydrolysis of the peptide on the carboxyl side of the asparagines shown Haemaphysalis longicornis Okayama ?
-
 ?
3.4.22.34 Hemoglobin + H2O strict specificity for hydrolysis of the peptide on the carboxyl side of the asparagines indicated Haemaphysalis longicornis ?
-
 ?
3.4.22.34 Hemoglobin + H2O strict specificity for hydrolysis of the peptide on the carboxyl side of the asparagines indicated Haemaphysalis longicornis Okayama ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.22.34 Asparaginyl endopeptidase
-
 Haemaphysalis longicornis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.22.34 37
-
 activity assay at Haemaphysalis longicornis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.4.22.34 10 70 assay under different temperature conditions tested Haemaphysalis longicornis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.22.34 7
-
 activity assay at Haemaphysalis longicornis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.22.34 3 11 assay under different pH conditions tested Haemaphysalis longicornis