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Literature summary extracted from
- HlLgm2, a member of asparaginyl endopeptidases/legumains in the midgut of the ixodid tick Haemaphysalis longicornis, is involved in blood-meal digestion (2008), J. Insect Physiol., 54, 573-585.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.22.34 | additional information | endogenous HlLgm2 expression in the midgut that is upregulated by the blood-feeding process is shown in all the developmental stages | Haemaphysalis longicornis |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.4.22.34 | expressed in Escherichia coli | Haemaphysalis longicornis |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.22.34 | antipain | papain-like cysteine proteinase inhibitor, 28.77% inhibition at 0.1 mM concentration relative to control | Haemaphysalis longicornis |  |
| 3.4.22.34 | Ca2+ | moderate inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | Co2+ | strong inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | Cu2+ | strong inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | cystatin | from egg white, 62.3% inhibition at 0.003 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | E-64 | papain-like cysteine proteinase inhibitor, 7.51% inhibition at 0.3 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | Fe2+ | strong inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | iodoacetamide | 100% inhibition at 3 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | leupeptin | papain-like cysteine proteinase inhibitor, 3.9% inhibition at 0.3 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | Mg2+ | moderate inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | Mn2+ | moderate inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | N-ethylmaleimide | 100% inhibition at 3 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | Ni2+ | strong inhibition | Haemaphysalis longicornis | |
| 3.4.22.34 | pepstatin A | aspartic peptidase inhibitor has minimal effects on the enzyme activity, 12.22% inhibition at 0.3 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | PMSF | serine peptidase inhibitor, 13.84% inhibition at 5 mM concentration relative to control | Haemaphysalis longicornis | |
| 3.4.22.34 | Zn2+ | moderate inhibition | Haemaphysalis longicornis | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.4.22.34 | 38000 | - |
endogenous HlLgm2, predicted and confirmed by SDS-PAGE and immunoblot analysis | Haemaphysalis longicornis |
| 3.4.22.34 | 42500 | - |
His-tagged rHlLgm2, determined by SDS-PAGE | Haemaphysalis longicornis |
| 3.4.22.34 | 47490 | - |
predicted preprotein | Haemaphysalis longicornis |
| 3.4.22.34 | 49630 | - |
predicted preproprotein | Haemaphysalis longicornis |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.22.34 | bovine albumin + H2O | Haemaphysalis longicornis | - |
peptides containing C-terminal Asn | - |
? | |
| 3.4.22.34 | bovine haemoglobin + H2O | Haemaphysalis longicornis | exhibited strict specificity for the asparaginyl bonds on the carboxy-terminal side of a peptide | peptides containing C-terminal Asn | - |
? | |
| 3.4.22.34 | additional information | Haemaphysalis longicornis | enzyme is involved in host blood-meal digestion | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.4.22.34 | Haemaphysalis longicornis | A9CQC1 | - |
- |
Posttranslational Modification
| EC Number | Posttranslational Modification | Comment | Organism |
|---|---|---|---|
| 3.4.22.34 | glycoprotein | asparagine-linked glycosylation | Haemaphysalis longicornis |
| 3.4.22.34 | proteolytic modification | the values expected on the basis of the predicted MW indicating that post-translational processing of HlLgm2 occurred during biosynthesis to form a mature protein through removal of 9 kDa peptides from the precursor protein, putative C-terminal autocleavage at site Asn364-365 | Haemaphysalis longicornis |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.4.22.34 | recombinant protein is purified using Ni-Sepharos under native conditions | Haemaphysalis longicornis |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 3.4.22.34 | adult | - |
Haemaphysalis longicornis | - |
| 3.4.22.34 | larva | - |
Haemaphysalis longicornis | - |
| 3.4.22.34 | midgut | - |
Haemaphysalis longicornis | - |
| 3.4.22.34 | nymph | - |
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 | 0.000133 | - |
hydrolysis of substrate benzyloxycarbonyl-Ala-Ala-Asn-4-methyl-coumaryl-7-amide | Haemaphysalis longicornis |
| 3.4.22.34 | 0.00041 | - |
hydrolysis of substrate acetyl-aspartyl-asparaginyl-leucyl-aspartic acid alpha-(4-methylcoumaryl-7-amide) | Haemaphysalis longicornis |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.22.34 | Ac-Asp-Asn-Leu-Asp alpha-(4-methylcoumaryl-7-amide) + H2O | - |
Haemaphysalis longicornis | Ac-Asp-Asn-Leu-Asp + 7-amino-4-methylcoumarin | - |
? | |
| 3.4.22.34 | benzyloxycarbonyl-Ala-Ala-Asn-7-amido-4-methylcoumarin + H2O | - |
Haemaphysalis longicornis | benzyloxycarbonyl-Ala-Ala-Asn + 7-amino-4-methylcoumarin | - |
? | |
| 3.4.22.34 | bovine albumin + H2O | - |
Haemaphysalis longicornis | peptides containing C-terminal Asn | - |
? | |
| 3.4.22.34 | bovine haemoglobin + H2O | exhibited strict specificity for the asparaginyl bonds on the carboxy-terminal side of a peptide | Haemaphysalis longicornis | peptides containing C-terminal Asn | - |
? | |
| 3.4.22.34 | additional information | enzyme is involved in host blood-meal digestion | Haemaphysalis longicornis | ? | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.4.22.34 | Asparaginyl endopeptidase | - |
Haemaphysalis longicornis |
| 3.4.22.34 | HlLgm2 | - |
Haemaphysalis longicornis |
| 3.4.22.34 | legumain | - |
Haemaphysalis longicornis |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.22.34 | 37 | - |
- |
Haemaphysalis longicornis |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.22.34 | 7 | - |
the rHlLgm2 is more active in neutral to slightly alkaline condition | Haemaphysalis longicornis |
pI Value
| EC Number | Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|---|
| 3.4.22.34 | Haemaphysalis longicornis | predicted preprotein | - |
6.7 |
| 3.4.22.34 | Haemaphysalis longicornis | predicted preproprotein | - |
7.1 |
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