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Literature summary extracted from

  • Martinez, M.; Diaz-Mendoza, M.; Carrillo, L.; Diaz, I.
    Carboxy terminal extended phytocystatins are bifunctional inhibitors of papain and legumain cysteine proteinases (2007), FEBS Lett., 581, 2914-2918.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.22.2 barley cystatin protease inhibitor inhibited by various cystatin vatiants, HvCPI-1, HvCPI-2, HvCPI-3, HvCPI-4, HvCPI-5, HvCPI-6 Carica papaya
3.4.22.2 additional information not inhibited by barley cystatin protease inhibitor-4 (Q86P), barley cystatin protease inhibitor-4 (N-term-DELTAA142 (Q86P)), barley cystatin protease inhibitor-4 (N-term-DELTAL150 (Q86P)), barley cystatin protease inhibitor-4 (DELTAT143-C-term), barley cystatin protease inhibitor-4 (DELTAT143-C-term (N177K)), barley cystatin protease inhibitor-4 (DELTAG151-C-term), and barley cystatin protease inhibitor-4 (DELTAG151-C-term (N177K)) Carica papaya
3.4.22.34 FaCPI-1 strawberry cystatin, phytocystatin of Fragaria sp. Hordeum vulgare
3.4.22.34 HvCPI-4 cystatin HvCPI-4 of Hordeum vulgare Hordeum vulgare
3.4.22.34 HvCPI-4-deltaT143-Cterm C-terminal extended part of cystatin HvCPI-4 Hordeum vulgare
3.4.22.34 HvCPI-4-deltaT151-Cterm C-terminal extended part of cystatin HvCPI-4 Hordeum vulgare
3.4.22.34 HvCPI-4-Q86P cystatin HvCPI-4 of Hordeum vulgare, mutant Q86P Hordeum vulgare
3.4.22.34 additional information inhibitory activity of the phytocystatins HvCPI-4 of Hordeum vulgare and FaCPI-1 of Fragaria sp. measured by the inhibition of legumain proteinase activity, ability of HvCPI-4 and FaCPI-1 to inhibit human legumain in vitro demonstrated Homo sapiens
3.4.22.34 additional information inhibitory activity measured by the inhibition of legumain proteinase activity, phytocystatins that are able to inhibit human legumain also inhibit the barley legumain proteinase activities in developing roots, endogenous barley legumain activities inhibited by iodoacetamide, no inhibition by the non-legumain inhibitors leupeptin and E64 observed; inhibitory effects against endogenous legumain activity in barley protein extracts tested, inhibitory activity measured by the inhibition of legumain-like proteinase activity, phytocystatins able to inhibit human legumain also inhibit the barley legumain proteinase activities in developing endosperms, endogenous barley legumain activities inhibited by iodoacetamide, no inhibition by the non-legumain inhibitors leupeptin and E64 observed Hordeum vulgare

Organism

EC Number Organism UniProt Comment Textmining
3.4.22.2 Carica papaya
-
-
-
3.4.22.34 Homo sapiens Q99538 Homo sapiens
-
3.4.22.34 Hordeum vulgare
-
 legumain activity in barley protein extracts analyzed
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.4.22.34 commercial preparation inhibitory effects by phytocystatins against commercial human legumain analyzed Homo sapiens
-
3.4.22.34 endosperm developing endosperms of 18 days after flowering, inhibitory effects by cystatins against endogenous legumain activity in barley protein extracts analyzed Hordeum vulgare
-
3.4.22.34 root collected from plants germinated at 22°C in the dark for 7 days Hordeum vulgare
-

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 additional information
-
 ability of carboxy-terminal extensions and mutants of phytocystatin to inhibit endogenous legumain activity present in barley protein extracts analyzed, inhibitory activity measured by the inhibition of legumain proteinase activity, assay described Hordeum vulgare
3.4.22.34 additional information
-
 ability of carboxy-terminal extensions and mutants of phytocystatin to inhibit in vitro human legumain shown, inhibitory activity measured by the inhibition of legumain proteinase activity, assay described Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.22.2 N-benzoyl-DL-arginine-2-naphthylamide + H2O
-
 Carica papaya N-benzoyl-DL-arginine + 2-naphthylamine
-
 ?
3.4.22.34 benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin-7-amide + H2O inhibitory activity of cystatin variants of Hordeum vulgare against endogenous legumain activity in protein extracts of Hordeum vulgare analyzed Hordeum vulgare benzyloxycarbonyl-Ala-Ala-Asn + 7-amino-4-methylcoumarin
-
 ?
3.4.22.34 benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin-7-amide + H2O inhibitory activity of cystatin variants of Hordeum vulgare against endogenous legumain activity in protein extracts of roots of Hordeum vulgare analyzed Hordeum vulgare benzyloxycarbonyl-Ala-Ala-Asn + 7-amino-4-methylcoumarin
-
 ?
3.4.22.34 benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin-7-amide + H2O inhibitory activity of phytocystatin variants of Hordeum vulgare tested against commercial human legumain Homo sapiens benzyloxycarbonyl-Ala-Ala-Asn + 7-amino-4-methylcoumarin
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.4.22.34 AEP
-
 Hordeum vulgare
3.4.22.34 AEP
-
 Homo sapiens
3.4.22.34 asparagine endopeptidase
-
 Hordeum vulgare
3.4.22.34 asparagine endopeptidase
-
 Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.4.22.2 additional information
-
 additional information inhibition constants for various barley cystatin protease inhibitor variants Carica papaya
3.4.22.34 0.00013
-
 HvCPI-4-Q86P mutant Q86P of HvCPI-4, inhibitor concentrations between 25 and 500 nM used, incubation with 10 microgram of a soluble endosperm protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00013
-
 HvCPI-4-Q86P mutant Q86P of HvCPI-4, inhibitor concentrations between 25 and 500 nM used, incubation with 5 microgram of a soluble root protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00019
-
 HvCPI-4 inhibitor concentrations between 25 and 500 nM used, incubation with 10 microgram of a soluble endosperm protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00019
-
 HvCPI-4-deltaT143-Cterm inhibitor concentrations between 25 and 500 nM used, incubation with 10 microgram of a soluble endosperm protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00019
-
 HvCPI-4 inhibitor concentrations between 25 and 500 nM used, incubation with 5 microgram of a soluble root protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00019
-
 HvCPI-4-deltaT143-Cterm inhibitor concentrations between 25 and 500 nM used, incubation with 5 microgram of a soluble root protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00029
-
 HvCPI-4-deltaT151-Cterm inhibitor concentrations between 25 and 500 nM used, incubation with 10 microgram of a soluble endosperm protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00029
-
 HvCPI-4-deltaT151-Cterm inhibitor concentrations between 25 and 500 nM used, incubation with 5 microgram of a soluble root protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00076
-
 FaCPI-1 inhibitor concentrations between 25 and 500 nM used, incubation with 10 microgram of a soluble endosperm protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare
3.4.22.34 0.00076
-
 FaCPI-1 inhibitor concentrations between 25 and 500 nM used, incubation with 5 microgram of a soluble root protein extract, at 25°C and pH 6 for 10 min Hordeum vulgare

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.4.22.34 additional information
-
 inhibitory doses for 50% proteinase activity inhibition higher for root legumain activity than for endosperm one Hordeum vulgare additional information
3.4.22.34 0.018
-
 endosperm Hordeum vulgare HvCPI-4-deltaT143-Cterm
3.4.22.34 0.02
-
 endosperm Hordeum vulgare FaCPI-1
3.4.22.34 0.022
-
 endosperm Hordeum vulgare HvCPI-4-deltaT151-Cterm
3.4.22.34 0.033
-
 endosperm Hordeum vulgare HvCPI-4-Q86P
3.4.22.34 0.038
-
 endosperm Hordeum vulgare HvCPI-4
3.4.22.34 0.068
-
 root extract Hordeum vulgare HvCPI-4-deltaT143-Cterm
3.4.22.34 0.091
-
 root extract Hordeum vulgare HvCPI-4-deltaT151-Cterm
3.4.22.34 0.11
-
 root extract Hordeum vulgare HvCPI-4-Q86P
3.4.22.34 0.12
-
 root extract Hordeum vulgare HvCPI-4
3.4.22.34 0.2
-
 root extract Hordeum vulgare FaCPI-1