Information on EC 3.4.22.35 - Histolysain

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The expected taxonomic range for this enzyme is: Entamoeba

EC NUMBER
COMMENTARY
3.4.22.35
-
RECOMMENDED NAME
GeneOntology No.
Histolysain
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hydrolysis of proteins, including basement membrane collagen and azocasein. Preferential cleavage: Arg-Arg-/- in small molecule substrates including Z-Arg-Arg-/-NHMec
show the reaction diagram
; from the protozoan, Entamoeba histolytica. In peptidase family C1 (papain family)
-
-
-
hydrolysis of proteins, including basement membrane collagen and azocasein. Preferential cleavage: Arg-Arg-/- in small molecule substrates including Z-Arg-Arg-/-NHMec
show the reaction diagram
cleavage mechanism on mucin substrate, overview
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
hydrolysis of peptide bond
Entamoeba histolytica HM1:IMSS
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Amebapain
-
-
-
-
amoebapain
-
-
CP1
Entamoeba histolytica HM1:IMSS
-
isoform
-
CP2
Entamoeba histolytica HM1:IMSS
-
isoform
-
CP5
Entamoeba histolytica HM1:IMSS
-
isoform
-
cysteine protease
-
-
cysteine protease 1
Q10957
-
cysteine protease 1
Q10957
one of the three major cysteine proteases
cysteine protease 2
-
-
cysteine protease 2
Q10958
-
cysteine protease 2
Q10958
one of the three major cysteine proteases
cysteine protease 5
-
-
cysteine protease 5
-
one of the three major cysteine proteases
cysteine proteinase 1
-
-
cysteine proteinase 1
Entamoeba histolytica HM1:IMSS
-
-
-
cysteine proteinase 5
-
-
cysteine proteinase 5
Entamoeba histolytica HM1:IMSS
-
-
-
EhCP1
Entamoeba histolytica HM1:IMSS
-
-
-
Entamoeba histolytica 112kDa adhesin
-
together with a protein with an adherence domain EhADH112
Entamoeba histolytica cysteine protease
-
-
-
-
Entamoeba histolytica cysteine protease
-
-
Entamoeba histolytica cysteine protease
Entamoeba histolytica HM1:IMSS
-
-
-
Entamoeba histolytica cysteine proteinase
-
-
-
-
Entamoeba histolytica cysteine proteinase
-
-
Entamoeba histolytica cysteine proteinase 5
-
-
Entamoeba histolytica cysteine-proteinase
-
-
Entamoeba histolytica neutral thiol proteinase
-
-
-
-
Histolysin
-
-
-
-
PCP5
-
isoform
PCP5
Entamoeba histolytica HM1:IMSS
-
isoform
-
Proteinase, Entamoeba histolytica cysteine
-
-
-
-
additional information
-
the enzyme belongs to the C1 peptidase family
additional information
Q95030
enzyme is a member of the cysteine proteinase family
CAS REGISTRY NUMBER
COMMENTARY
92228-52-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
gene edcp2 or acp2
-
-
Manually annotated by BRENDA team
control strain and strains overexpressing EhRab7B and EhRab7B-GTP, activity secreted to the medium by the EhRab7B-GTP transformant is 4.3fold higher than the control transformant and has a 38% less total intracellular CP activity than the control transformant
-
-
Manually annotated by BRENDA team
enzyme is termed CP5
SwissProt
Manually annotated by BRENDA team
gene ehcp112
-
-
Manually annotated by BRENDA team
gene Ehcp112, gene is encoded adjacent to Ehadh112
-
-
Manually annotated by BRENDA team
gene ehcp2
-
-
Manually annotated by BRENDA team
gene ehcp2 or acp2 encodes histolysain, gene ehcp1 or acp1 encodes amoebapain
-
-
Manually annotated by BRENDA team
gene ehcp5
-
-
Manually annotated by BRENDA team
isolate HM-1:IMSS, genes ehcp1, ehcp2, and ehcp5
-
-
Manually annotated by BRENDA team
low-virulence HMI strain and highly virulent 1659 clone, derived from HMI by hamster liver passages
-
-
Manually annotated by BRENDA team
overexpression of EhRab11B protein results in a 15fold increase in both, intracellular and secreted cysteine protease activity, suggesting that the three major cysteine proteases are trafficked via an EhRab11B-associated secretory pathway
-
-
Manually annotated by BRENDA team
overexpression of EhRab11B protein results in a 15fold increase in both, intracellular and secreted cysteine protease activity, suggesting that the three major cysteine proteases are trafficked via an EhRab11B-associated secretory pathway
UniProt
Manually annotated by BRENDA team
strain HM 1:IMSS, catalytic classes of proteinases
-
-
Manually annotated by BRENDA team
strain HM1:IMSS
-
-
Manually annotated by BRENDA team
strain HM1:IMSS and its derivative clone A
-
-
Manually annotated by BRENDA team
virulent strain HMI:IMSS
-
-
Manually annotated by BRENDA team
Entamoeba histolytica HM 1:IMSS
strain HM 1:IMSS, catalytic classes of proteinases
-
-
Manually annotated by BRENDA team
Entamoeba histolytica HM1:IMSS
-
-
-
Manually annotated by BRENDA team
Entamoeba histolytica HM1:IMSS
strain HM1:IMSS
-
-
Manually annotated by BRENDA team
Entamoeba histolytica HM1:IMSS
strain HM1:IMSS and its derivative clone A
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
cysteine proteinase 5 is a major virulent factor that binds integrin on colonic cells and stimulates nuclear factor kappaB-mediated pro-inflammatory responses. Cysteine proteinase 5 is the critical proteinase involved in activating nuclear factor kappaB, phosphorylating Akt and inducing the ubiquitination of nuclear factor-kappaB essential modulator
physiological function
Entamoeba histolytica HM1:IMSS
-
cysteine proteinase 5 is a major virulent factor that binds integrin on colonic cells and stimulates nuclear factor kappaB-mediated pro-inflammatory responses. Cysteine proteinase 5 is the critical proteinase involved in activating nuclear factor kappaB, phosphorylating Akt and inducing the ubiquitination of nuclear factor-kappaB essential modulator
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Arg-Gly-Phe-Phe + H2O
Arg-Gly + Phe-Phe
show the reaction diagram
-
substrate of amoebapain
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
-
-
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
?
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
?
Azocoll + H2O
?
show the reaction diagram
-
-
-
-
?
Azocoll + H2O
Hydrolyzed azocoll
show the reaction diagram
-
-
-
-
-
benzyloxycarbonyl-Ala-Arg-Arg-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Arg-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
Benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
benzyloxycarbonyl-Arg-Arg-4-nitroanilide + H2O
benzyloxycarbonyl-Arg-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Arg-Arg-4-nitroanilide + H2O
benzyloxycarbonyl-Arg-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
Q10957, Q10958
-
-
-
?
benzyloxycarbonyl-L-arginyl-L-arginine 4-nitroanilide + H2O
benzyloxycarbonyl-L-arginyl-L-arginine + 4-nitroaniline
show the reaction diagram
Q95030
-
-
?
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Phe-L-citrullin 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
?
C3 protein + H2O
?
show the reaction diagram
-
protein from human
-
-
?
Cartilage proteoglycan + H2O
?
show the reaction diagram
-
-
-
-
-
chemokine CCL13 + H2O
?
show the reaction diagram
-
-, Entamoeba histolytica is a human pathogen causing amoebic colitis, the enzyme is involved in the inflammation process modulating human host leucocyte migration by proteolytic cleavage of chemokines CCL2, CCL13, and CXCL8, a mechanism to circumvent host immune response
-
-
?
chemokine CCL2 + H2O
?
show the reaction diagram
-
-, Entamoeba histolytica is a human pathogen causing amoebic colitis, the enzyme is involved in the inflammation process modulating human host leucocyte migration by proteolytic cleavage of chemokines CCL2, CCL13, and CXCL8, a mechanism to circumvent host immune response
-
-
?
chemokine CXCL8 + H2O
?
show the reaction diagram
-
-, Entamoeba histolytica is a human pathogen causing amoebic colitis, the enzyme is involved in the inflammation process modulating human host leucocyte migration by proteolytic cleavage of chemokines CCL2, CCL13, and CXCL8, a mechanism to circumvent host immune response
-
-
?
Collagen + H2O
?
show the reaction diagram
-
-
-
?
collagen type 1 + H2O
?
show the reaction diagram
-
-
-
-
?
Collagen type I + H2O
?
show the reaction diagram
-
digestion with an initial attack at the alpha2-chain
-
-
-
complement factor C3 + H2O
peptides
show the reaction diagram
Q95030
-
product determination
?
complement factor C9 + H2O
peptides
show the reaction diagram
Q95030
-
product determination
?
Fibronectin + H2O
?
show the reaction diagram
-
-
-
?
Fibronectin + H2O
?
show the reaction diagram
-
-
-
-
?
fibronectin + H2O
peptides
show the reaction diagram
Q95030
-
product determination
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
Entamoeba histolytica HM1:IMSS
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
?
hide powder azure + H2O
?
show the reaction diagram
-
-
-
-
-
immunoglobulin G + H2O
peptides derived from immunoglobulin G
show the reaction diagram
Q95030
-
product determination
?
immunoglobulin G + H2O
?
show the reaction diagram
-
-
-
?
Muc2 + H2O
?
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
-
-
-
?
MUC2 mucin + H2O
?
show the reaction diagram
-
disruption of the protective mucin layer of targeted host cells, entry mechanism of the pathogen, overview, the enzyme targets two site at the C-terminal cysteine-rich domain, which is less glycosylated and exposes its peptide chain, overview
-
-
?
mucin + H2O
?
show the reaction diagram
-
-
-
?
mucin + H2O
?
show the reaction diagram
-
-
-
?
mucin MUC2 polymer + H2O
?
show the reaction diagram
-
enzyme is involved, together with other Entamoeba histolytica cysteine proteinases, in the degradation of mucin in the mucous layer of the colon epithelium, altering the protective function to facilitate parasite attachment
-
?
N-benzyloxycarbonyl-L-arginyl-7-amido-4-methylcoumarin + H2O
N-benzyloxycarbonyl-L-arginine + 7-amino-4-methylcoumarin
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
17% of the activity with N-benzyloxycarbonyl-L-arginyl-L-arginyl-7-amido-4-methylcoumarin
-
?
N-benzyloxycarbonyl-L-arginyl-L-arginyl-7-amido-4-methylcoumarin + H2O
N-benzyloxycarbonyl-L-arginyl-L-arginine + 7-amino-4-methylcoumarin
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
-
-
?
N-benzyloxycarbonyl-L-phenylalanyl-L-arginyl-7-amido-4-methylcoumarin + H2O
N-benzyloxycarbonyl-L-phenylalanyl-L-arginine + 7-amino-4-methylcoumarin
show the reaction diagram
-
7% of the activity with N-benzyloxycarbonyl-L-arginyl-L-arginyl-7-amido-4-methylcoumarin
-
?
pro-interleukin-18 + H2O
?
show the reaction diagram
-
protein from human
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
Q95030
enzyme is involved in pathogenic destruction of host tissue by degradation of extracellular matrix proteins
-
?
protein + H2O
peptides
show the reaction diagram
Entamoeba histolytica HM1:IMSS
-
-
-
?
Z-Ala-Arg-Arg-7-amido-4-methylcoumarin + H2O
Z-Ala-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
-
-
-
?
Z-Arg-Arg-4-methoxy-2-nitroanilide + H2O
Z-Arg-Arg + 4-methoxy-2-nitroaniline
show the reaction diagram
-
-
-
-
?
Z-Arg-Arg-4-nitroanilide + H2O
Z-Arg-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Z-Arg-Arg-7-amido-4-methylcoumarin + H2O
Z-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
-
-
-
?
Z-Arg-Arg-7-amido-4-trifluoromethylcoumarin + H2O
Z-Arg-Arg + 7-amino-4-trifluoromethylcoumarin
show the reaction diagram
-, Q10957, Q10958
-
-
-
?
Z-Arg-Arg-7-amido-4-trifluoromethylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
immunoglobulin G + H2O
?
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
heavy chain, protein from human
-
-
?
Insulin B-chain + H2O
additional information
-
-
-
the Gly-Phe bond in the insulin B-chain is the major hydrolysis site
-
Kidney glomerular basement-membrane collagen + H2O
?
show the reaction diagram
-
-
-
-
-
additional information
?
-
-
with unblocked tetrapeptides as substrates, peptidyl dipeptidase activity of the amoeba enzyme requires an arginine at the P2 position. Lysine cannot substitute for arginine
-
-
-
additional information
?
-
-
the enzyme causes a loss of adhesion of mammalian cells in culture, plasminogen activator
-
-
-
additional information
?
-
-
elastin
-
-
-
additional information
?
-
-
inactivates aldolase and glyceraldehyde 3-phosphate dehydrogenase from rabbit muscle and glucose 6-phosphate dehydrogenase from yeast, limited proteolysis yielding major cleavage products
-
-
-
additional information
?
-
-
not: type I collagen
-
-
-
additional information
?
-
-
splits blocked and unblocked peptide analogs with 2-naphthylamide moieties, cleavability is enhanced by the presence of basic residues, such as arginine or lysine, near the acyl end of the substrate
-
-
-
additional information
?
-
-
enzyme permits adhesion of trophozoites to cells via fibronectin binding, which can be inhibited by Zn2+
-
?
additional information
?
-
-
no activity with benzyloxycarbonyl-Phe-Arg-4-nitroanilide
-
?
additional information
?
-
-
no activity with N-benzyloxycarbonyl-Val-Lys-Met-7-amido-4-methylcoumarin and succinyl-LLVY-7-amido-4-methylcoumarin
-
?
additional information
?
-
Q95030
no activity with peptide substrates containing phenylalanine at P2 position instead of arginine
-
?
additional information
?
-
-
the enzyme is covalently connected to an adhesin by electrostatic forces which are not broken during phagocytosis
-
?
additional information
?
-
-
significant pathogenicity factor
-
-
-
additional information
?
-
-
role in tissue invasion
-
-
-
additional information
?
-
-
appears to be important both for digestion and as a cytotoxic factor
-
-
-
additional information
?
-
-
the enzyme can degrade colon cell mucin and destroy epithelial cell layers
-
-
-
additional information
?
-
-
the enzyme degrades a number of proteins, including those of the extracellular matrix
-
-
-
additional information
?
-
-
the enzyme is involved in host tissue invasion by trophozoites and destroys cell monolayers
-
-
-
additional information
?
-
-
the enzyme shows cytopathic effects and destroys cell monolayers
-
-
-
additional information
?
-
-
the enzyme degrades a number of proteins, including those of the extracellular matrix, but also acts on small molecule substrates showing endopeptidase and exopeptidase-like activities, amoebapain hydrolyzes unblocked tetrapeptides with basic residues at P2
-
-
-
additional information
?
-
-
cysteine proteinases play a central role in tissue invasion and disruption of host defenses by digesting components of the extracellular matrix, immunoglobulins, complement, and cytokines
-
-
-
additional information
?
-
-
essential for pathogenicity of Entamoeba histolytica
-
-
-
additional information
?
-
Q10957, Q10958
essential for pathogenicity of Entamoeba histolytica
-
-
-
additional information
?
-
-, Q10957, Q10958
secreted cysteine proteases play an indispensable role in amoebic invasion and tissue destruction due to their hydrolytic and degradative activities towards extracellular matrix proteins
-
-
-
additional information
?
-
-
no activity with Z-Phe-Arg-4-amido-7-methylcoumarin
-
-
-
additional information
?
-
-
the cysteine proteinase EhCP112 and the adhesin EhADH112 assemble to form the EhCPADH complex involved in Entamoeba histolytica virulence
-
-
-
additional information
?
-
Entamoeba histolytica HM1:IMSS
-
cysteine proteinases play a central role in tissue invasion and disruption of host defenses by digesting components of the extracellular matrix, immunoglobulins, complement, and cytokines, no activity with Z-Phe-Arg-4-amido-7-methylcoumarin
-
-
-
additional information
?
-
Entamoeba histolytica HM1:IMSS
-
no activity with N-benzyloxycarbonyl-Val-Lys-Met-7-amido-4-methylcoumarin and succinyl-LLVY-7-amido-4-methylcoumarin
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
chemokine CCL13 + H2O
?
show the reaction diagram
-
Entamoeba histolytica is a human pathogen causing amoebic colitis, the enzyme is involved in the inflammation process modulating human host leucocyte migration by proteolytic cleavage of chemokines CCL2, CCL13, and CXCL8, a mechanism to circumvent host immune response
-
-
?
chemokine CCL2 + H2O
?
show the reaction diagram
-
Entamoeba histolytica is a human pathogen causing amoebic colitis, the enzyme is involved in the inflammation process modulating human host leucocyte migration by proteolytic cleavage of chemokines CCL2, CCL13, and CXCL8, a mechanism to circumvent host immune response
-
-
?
chemokine CXCL8 + H2O
?
show the reaction diagram
-
Entamoeba histolytica is a human pathogen causing amoebic colitis, the enzyme is involved in the inflammation process modulating human host leucocyte migration by proteolytic cleavage of chemokines CCL2, CCL13, and CXCL8, a mechanism to circumvent host immune response
-
-
?
collagen type 1 + H2O
?
show the reaction diagram
-
-
-
-
?
Muc2 + H2O
?
show the reaction diagram
Entamoeba histolytica, Entamoeba histolytica HM1:IMSS
-
-
-
-
?
MUC2 mucin + H2O
?
show the reaction diagram
-
disruption of the protective mucin layer of targeted host cells, entry mechanism of the pathogen, overview
-
-
?
mucin MUC2 polymer + H2O
?
show the reaction diagram
-
enzyme is involved, together with other Entamoeba histolytica cysteine proteinases, in the degradation of mucin in the mucous layer of the colon epithelium, altering the protective function to facilitate parasite attachment
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
Q95030
enzyme is involved in pathogenic destruction of host tissue by degradation of extracellular matrix proteins
-
?
protein + H2O
peptides
show the reaction diagram
Entamoeba histolytica HM1:IMSS
-
-
-
?
Fibronectin + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
significant pathogenicity factor
-
-
-
additional information
?
-
-
role in tissue invasion
-
-
-
additional information
?
-
-
appears to be important both for digestion and as a cytotoxic factor
-
-
-
additional information
?
-
-
the enzyme can degrade colon cell mucin and destroy epithelial cell layers
-
-
-
additional information
?
-
-
the enzyme degrades a number of proteins, including those of the extracellular matrix
-
-
-
additional information
?
-
-
the enzyme is involved in host tissue invasion by trophozoites and destroys cell monolayers
-
-
-
additional information
?
-
-
the enzyme shows cytopathic effects and destroys cell monolayers
-
-
-
additional information
?
-
-
cysteine proteinases play a central role in tissue invasion and disruption of host defenses by digesting components of the extracellular matrix, immunoglobulins, complement, and cytokines
-
-
-
additional information
?
-
-
essential for pathogenicity of Entamoeba histolytica
-
-
-
additional information
?
-
Q10957, Q10958
essential for pathogenicity of Entamoeba histolytica
-
-
-
additional information
?
-
-, Q10957, Q10958
secreted cysteine proteases play an indispensable role in amoebic invasion and tissue destruction due to their hydrolytic and degradative activities towards extracellular matrix proteins
-
-
-
additional information
?
-
-
the cysteine proteinase EhCP112 and the adhesin EhADH112 assemble to form the EhCPADH complex involved in Entamoeba histolytica virulence
-
-
-
additional information
?
-
Entamoeba histolytica HM1:IMSS
-
cysteine proteinases play a central role in tissue invasion and disruption of host defenses by digesting components of the extracellular matrix, immunoglobulins, complement, and cytokines
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4-hydroxymercuribenzoate
-
E64 inhibits cell growth and is cytopathic/lethal for E64-sensitive and E64-resistant amoebae
4-[(2S,3S)-carboxyoxiran-2-ylcarbonyl-L-leucylamido]butylguanidine
-
i.e. trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane or E64
Chicken cystatin
-
-
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Cystatin
-
-
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cysteine protease inhibitor 1
-
-
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cysteine protease inhibitor 2
-
inhibits isoform CP5 approximately 20times more efficient than isoform CP2
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E64
-
specific inhibition
E64
-, Q10957, Q10958
activity was completely abolished when trophozoites are pretreated with 200 mM of E64, a potent, irreversible and highly selective cysteine protease inhibitor; activity was completely abolished when trophozoites are pretreated with 200 mM of E64, a potent, irreversible and highly selective cysteine protease inhibitor; activity was completely abolished when trophozoites are pretreated with 200 mM of E64, a potent, irreversible and highly selective cysteine protease inhibitor
EDTA
-
slight inhibition at 10 mM
EhICP1
Q10957, Q10958
Entamoeba histolytica inhibitor for cysteine protease type 1; Entamoeba histolytica inhibitor for cysteine protease type 1; Entamoeba histolytica inhibitor for cysteine protease type 1, inhibition also by regulation of CP5 processing
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EhICP2
Q10957, Q10958
Entamoeba histolytica inhibitor for cysteine protease type 2; Entamoeba histolytica inhibitor for cysteine protease type 2; Entamoeba histolytica inhibitor for cysteine protease type 2, inhibition also by regulation of CP5 processing
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iodoacetic acid
-
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L-3-carboxy-2,3-trans-epoxypropionyl-leucyl-amido(4-guanidino)butane
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i.e. E-64
L-trans-epoxysuccinyl-(ethoxy)-Leu-3-methylbutylamide ethyl ester
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i.e. E64d, inhibits cell growth and is cytopathic/lethal for E64-sensitive and E64-resistant amoebae
L-trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
-
L-trans-epoxysuccinyl-Leu-4-guanidinobutylamide
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i.e. E64, inhibits cell growth in E64-resistant by about 80% and is cytopathic/lethal for E64-sensitive amoebae
N-ethylmaleimide
-
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N-methylpiperazine-urea-phenylalanylhomoarginylvinylsulfone-benzene
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WRR483, inhibitor addition to medium fully prevents invasion of parasite
N-methylpiperazine-urea-phenylalanylhomophenylalanylvinylsulfone-benzene
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K11777, inhibitor addition to medium partially prevents invasion of parasite
Nalpha-4-tosyl-L-lysine chloromethylketone
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Natural and synthetic inhibitors of cysteine proteinases
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-
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p-hydroxymercuribenzoate
Q95030
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Peptidyldiazomethanes
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Phenanthroline
-
slight inhibition at 10 mM
tosylsulfonylleucylchloromethyl ketone
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i.e. TLCK
tosylsulfonylphenylalanylchloromethyl ketone
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i.e. TPCK
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
-
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
Q95030
i.e. E64
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
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ie.e. E-64, irreversible
Z-Phe-Ala-diazomethylketone
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E64 inhibits cell growth and is cytopathic/lethal for E64-sensitive and E64-resistant amoebae
Z-Phe-Phe-diazomethylketone
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25% inhibition of E64-sensitive and E64-resistant amoba growth
Zn2+
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77% inhibition of proteolytic activity at 2.5 mM, completely reversible by L-cystein, phenanthroline, and L-histidine, Zn2+ interferes with the fibronectin binding of the amoeba
additional information
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no inhibition by pepstatin and phenylmethylsulfonyl fluoride
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additional information
-
no inhibition by aprotinin, pepstatin, phosphoramidon, and pefabloc SC
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ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
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stimulates
cysteine
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5-10 mM activate refolded protein
dithiothreitol
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activates
dithiothreitol
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2-5 mM activate refolded protein in a Tris, phosphate or citrate buffer containing EDTA, pH 7.0-7.5
sulfhydryl compounds
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activate
Liposomes
Q95030
association increases activity about 2fold
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additional information
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expression of ehpc1 and ehpc2 is increased in presence of E64 in E64-sensitive and E64-resistant amoebae, overview
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KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0015
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benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
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-
0.032
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Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
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TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
130
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benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
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-
0.4
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Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
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-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.132
-
cysteine protease inhibitor 1
-
isoform CP1, pH and temperature not specified in the publication
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0.59
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cysteine protease inhibitor 1
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isoform CP5, pH and temperature not specified in the publication
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0.809
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cysteine protease inhibitor 1
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isoform CP2, pH and temperature not specified in the publication
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0.05
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cysteine protease inhibitor 2
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isoform CP5, pH and temperature not specified in the publication
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0.166
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cysteine protease inhibitor 2
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isoform CP1, pH and temperature not specified in the publication
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0.95
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cysteine protease inhibitor 2
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isoform CP2, pH and temperature not specified in the publication
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1.3e-07
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EhICP1
Q10957, Q10958
pH 5.0, 37C
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5.9e-07
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EhICP1
Q10957, Q10958
pH 5.0, 37C
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8.1e-07
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EhICP1
Q10957, Q10958
pH 5.0, 37C
-
5e-08
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EhICP2
Q10957, Q10958
pH 5.0, 37C
-
1.7e-07
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EhICP2
Q10957, Q10958
pH 5.0, 37C
-
9.5e-07
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EhICP2
Q10957, Q10958
pH 5.0, 37C
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SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
170
-
-
purified native enzyme, substrate azocasein
180
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-
purified recombinant enzyme, substrate azocasein
470
-
Q95030
purified enzyme
890
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Q95030
purified enzyme associated to liposomes
1450
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-
refolded, recombinant, activated enzyme
2800
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purified enzyme
additional information
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-
-
additional information
-
-
cytopathic activity of wild-type and recombinant amoebae
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.4
-
-
azocasein
5
6
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dependent on the substrate
5.5
8
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recombinant enzyme
5.5
-
-
azocasein
6
-
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almost fully active between pH 5.5 and 7.5
6.7
-
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hide powder azure
7
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assay at, substrate azocasein
7.4
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-
assay at
7.5
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benzyloxycarbonyl-Phe-L-citrullin 4-methylcoumarin 7-amide, azocoll
9.5
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benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide, benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
additional information
-
-
neutral pH-optimum
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.4
8.5
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azocasein
4.5
9.5
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recombinant enzyme
5
8
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50% of activity at pH 5 and pH 8
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
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assay at, substrate azocasein
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
at optimal pH, the turnover increases with increasing temperature up to 85C
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.7
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isoelectric focusing
8.12
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Q95030
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additional information
-
-
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SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Entamoeba histolytica HM1:IMSS
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-
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Manually annotated by BRENDA team
additional information
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the enzyme is translocated to the amoebic surface upon the interaction of trophozoites with red blood cells, the enzyme participates thus in erythrophagocytosis
Manually annotated by BRENDA team
additional information
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the enzyme is expressed and active in different life cycle stages
Manually annotated by BRENDA team
additional information
Entamoeba histolytica HM1:IMSS
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the enzyme is translocated to the amoebic surface upon the interaction of trophozoites with red blood cells, the enzyme participates thus in erythrophagocytosis
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Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
the enzyme is secreted and localized on the cell surface
Manually annotated by BRENDA team
Entamoeba histolytica HM1:IMSS
-
-
-
Manually annotated by BRENDA team
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large cytoplasmic vesicles
Manually annotated by BRENDA team
Entamoeba histolytica HM1:IMSS
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large cytoplasmic vesicles
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Manually annotated by BRENDA team
Q95030
bound to the host cell surface
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Manually annotated by BRENDA team
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EhCP112 is secreted
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Manually annotated by BRENDA team
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the enzyme is secreted
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Manually annotated by BRENDA team
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lysosome-like vesicles
Manually annotated by BRENDA team
Q95030
associated by hydrophobic interaction
Manually annotated by BRENDA team
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the enzyme contains a transmembrane segment
Manually annotated by BRENDA team
additional information
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bound to matrix-like structures both on the cell surface and within subcellular vesicles. The enzyme recycles between plasma membrane and pinocytic vesicles
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Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
29000
-
-
Entamoeba histolytica, gel filtration
66000
-
-
native PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
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x * 56000, Entamoeba histolytica
?
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x * 112000, enzyme attached to adhesin, reducing and nonreducing SDS-PAGE, x * 75000, about, enzyme without adhesin, SDS-PAGE
?
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x * 27000-29000, recombinant and native enzyme, reducing and nonreducing SDS-PAGE
?
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x * 52000, recombinant His-tagged full-length EhCP112 zymogen, containing the signal peptide and the pro-peptide, SDS-PAGE
?
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x * 31000, SDS-PAGE, refolded, active protein; x * 50000, SDS-PAGE, predicted molecular mass for thio-pro-EhCP1 of 50300 Da (36300 Da for the zymogen form of EhCP1, 13000 Da for the thioredoxin peptide, and 9000 Da for the six-His tag)
?
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x * 52000, recombinant and refolded enzyme, SDS-PAGE, x * 35500, recombinant, refolded, and processed enzyme, SDS-PAGE
?
Entamoeba histolytica HM1:IMSS
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x * 31000, SDS-PAGE, refolded, active protein; x * 50000, SDS-PAGE, predicted molecular mass for thio-pro-EhCP1 of 50300 Da (36300 Da for the zymogen form of EhCP1, 13000 Da for the thioredoxin peptide, and 9000 Da for the six-His tag)
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monomer
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1 * 26000, Entamoeba histolytica, SDS-PAGE
monomer
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1 * 27000, Entamoeba histolytica, SDS-PAGE
monomer
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1 * 50000-60000, 3 forms, SDS-PAGE
monomer
-
1 * 27000, about
monomer
Entamoeba histolytica HM1:IMSS
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1 * 50000-60000, 3 forms, SDS-PAGE
-
additional information
Q95030
three-dimensional structure modeling
additional information
-
enzyme contains a transmembranal segment, amino acid residues 259-280, and an RGD motif
additional information
-
the enzyme contains an RDG domain
additional information
-
the enzyme contains an RGD integrin attachment motif, structure-function overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no glycoprotein
-
no N-glycosylation
proteolytic modification
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the recombinant enzyme needs to be refolded and processed from the inactive precursor, ppEhCP112, to a 35.5 kDa mature and active enzyme, EhCP112. The thiol specific inhibitor E-64, but not serine or aspartic proteinase inhibitors arrests this activation process
additional information
-
processing of the inactive pro-enzyme to the mature active form by treatment with 10 mM DTT and 0.04% SDS at 37C and pH 8.8, SDS can be substituted by Triton X-100 and octylglucoside but these are 3fold less efficient
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
9.5
-
4C, stable during overnight incubation
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
pH 5.5-9.5, stable during overnight incubation
additional information
-
-
Hg2+, at low concentration causes a delay in heat denaturation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Hg2+, at low concentration causes a delay in heat denaturation
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
15fold, to homogeneity
-
recombinant His-tagged enzyme from Escherichia coli
-
recombinant His-tagged full-length EhCP112 zymogen, containing the signal peptide and the pro-peptide, from Escherichia coli strain BL21 by nickel affinity chromatography
-
recombinant His-tagged inactive pro-enzyme to homogeneity
-
recombinant protein using His-tag under denaturating conditions
-
recombinant protein using His-tag under denaturating conditions; recombinant protein using His-tag under denaturating conditions; recombinant protein using His-tag under denaturating conditions
Q10957, Q10958
recombinant refolded His-tagged enzyme by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
DNA and amino acid sequence determination
Q95030
expressed as His-tag and thioredoxin fusin protein in Escherichia coli
-
expressed as His-tag fusion protein in Escherichia coli BL21(DE3), formation of inclusion bodies; expressed as His-tag fusion protein in Escherichia coli BL21(DE3), formation of inclusion bodies; expressed as His-tag fusion protein in Escherichia coli BL21(DE3), formation of inclusion bodies
Q10957, Q10958
expression of partial gene ehcp1 in Escherichia coli as GST-fusion protein, expression of functional EhCP2 in insect cells using the baculovirus transfection system
-
expression of the His-tagged inactive pro-enzyme in Escherichia coli BL21(DE3)
-
functional expression of EhCP5
-
gene ehcp112, functional expression of the His-tagged full-length EhCP112 zymogen, containing the signal peptide and the pro-peptide, in Escherichia coli strain BL21
-
gene Ehcp112, gene is encoded adjacent to Ehadh112, the latter encoding an adhesin that is covalently connected to the enzyme by electrostatic forces, DNA sequence determination and analysis, expression as His-tagged enzyme in Escherichia coli BL21(DE3)
-
genes ehcp1, ehcp2, and ehcp5, stable episomal transfection, overexpression of EhCP5 highly increases activity of EhCP1, EhCP2, and EhCP5 in recombinant amoeba and cytopathic activity, ehile overexpression of EhCP1 or 2 enhances only the single actvity and moderately enhances cytopathic activity, overview
-
recombinant expression of the His-tagged enzyme in Escherichia coli inclusion bodies
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
CP5 expression is significantly higher in the Entamoeba histolytica strain EGG isolated from liver than in the other strains maintained in culture (5.6fold higher than strain 32, 4.1fold higher than strain HM1 and 3.0fold higher than cultured strain EGG)
-
when Entamoeba histolytica is grown in the presence of E-64, PCP5 levels are significantly increased
-
when Entamoeba histolytica is grown in the presence of E-64, PCP5 levels are significantly increased
Entamoeba histolytica HM1:IMSS
-
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
construction of EhCP5-deficient amoeba by antisense inhibition showing no activity on mucin from cells LS174T and HT-29F Cl.16E or against CHO cell layers, enzyme-deficient amoeba cannot overcome the mucus barrier to disrupt CHO cell monolayers, overview
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
dialysis against refolding buffer; dialysis against refolding buffer; dialysis against refolding buffer
Q10957, Q10958
recombinant His-tagged enzyme, solvation in 20 mM sodium phosphate, pH 7.4, 0.5 M NaCl, 10 mM imidazole, and 8 M urea, refolding and processing of the inactive precursor, ppEhCP112, to a 35.5 kDa mature and active enzyme, EhCP112
-
renaturation of purified inactive pro-enzyme by a 100fold dilution in a buffer containing reduced and oxidized thiols leading to a soluble inactive pro-enzyme
-