Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.22.B68 extracted from

  • He, C.; Nora, G.P.; Schneider, E.L.; Kerr, I.D.; Hansell, E.; Hirata, K.; Gonzalez, D.; Sajid, M.; Boyd, S.E.; Hruz, P.; Cobo, E.R.; Le, C.; Liu, W.T.; Eckmann, L.; Dorrestein, P.C.; Houpt, E.R.; Brinen, L.S.; Craik, C.S.; Roush, W.R.; McKerrow, J.; Reed, S.L.
    A novel Entamoeba histolytica cysteine proteinase, EhCP4, is key for invasive amebiasis and a therapeutic target (2010), J. Biol. Chem., 285, 18516-18527.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.B68

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 Condon Plus (DE3)-RIPL cells as as a fusion protein with bacterial thioredoxin A with the His tag at the amino terminus of the prodomain Entamoeba histolytica

Inhibitors

Inhibitors Comment Organism Structure
WRR605 CP4-specific inhibitor, synthesized based on the substrate specificity of CP4, inhibits the recombinant enzyme in vitro Entamoeba histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
 benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin recombinant enzyme, at pH 7.0, 37°C Entamoeba histolytica
0.031
-
 succinyl-LLVY-7-amido-4-methylcoumarin recombinant enzyme, at pH 7.0, 37°C Entamoeba histolytica
0.046
-
 benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin recombinant enzyme, at pH 8.0, 37°C Entamoeba histolytica
0.078
-
 benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin recombinant enzyme, at pH 4.0, 37°C Entamoeba histolytica

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
 Entamoeba histolytica 5737
-
endoplasmic reticulum from perinuclear region Entamoeba histolytica 5783
-
nucleus
-
 Entamoeba histolytica 5634
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23000
-
 calculated molecular mass of the mature enzyme Entamoeba histolytica
26000
-
 active recombinant CP4, SDS-PAGE Entamoeba histolytica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
complement component C3 + H2O Entamoeba histolytica
-
 ? the primary product is a fragment of 70000 Da, a size similar to that of complement component C3b which is further proteolytically degraded by CP4 ?
complement component C3 + H2O Entamoeba histolytica HM1-IMSS
-
 ? the primary product is a fragment of 70000 Da, a size similar to that of complement component C3b which is further proteolytically degraded by CP4 ?
immunglobulin A + H2O Entamoeba histolytica
-
 ?
-
 ?
immunglobulin G + H2O Entamoeba histolytica
-
 ?
-
 ?
Laminin-1 + H2O Entamoeba histolytica
-
 ?
-
 ?
Laminin-1 + H2O Entamoeba histolytica HM1-IMSS
-
 ?
-
 ?
pro-interleukin-18 + H2O Entamoeba histolytica
-
 ?
-
 ?
villin-1 + H2O Entamoeba histolytica
-
 ?
-
 ?

Organism

Organism UniProt Comment Textmining
Entamoeba histolytica C4LTT0
-
-
Entamoeba histolytica HM1-IMSS C4LTT0
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel affinity column chromatography Entamoeba histolytica

Renatured (Commentary)

Renatured (Comment) Organism
maximal active enzyme is obtained by reducing the denatured thioredoxin A-CP4 fusion protein with 25 mM DTT at 37°C for 1 h followed by rapid dilution of the protein into ice-cold refolding buffer (100 mM Tris-HCl pH 8.5, 100 mM NaCl, 20% (v/v) glycerol, 250 mM arginine-HCl, 2 mM EDTA, pH 8, 10 mM GSH, 1 mM L-GSSG disodium salt) Entamoeba histolytica

Source Tissue

Source Tissue Comment Organism Textmining
trophozoite
-
 Entamoeba histolytica
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin + H2O
-
 Entamoeba histolytica benzyloxycarbonyl-VVR + 7-amino-4-methylcoumarin
-
 ?
complement component C3 + H2O
-
 Entamoeba histolytica ? the primary product is a fragment of 70000 Da, a size similar to that of complement component C3b which is further proteolytically degraded by CP4 ?
complement component C3 + H2O
-
 Entamoeba histolytica HM1-IMSS ? the primary product is a fragment of 70000 Da, a size similar to that of complement component C3b which is further proteolytically degraded by CP4 ?
immunglobulin A + H2O
-
 Entamoeba histolytica ?
-
 ?
immunglobulin G + H2O
-
 Entamoeba histolytica ?
-
 ?
Laminin-1 + H2O
-
 Entamoeba histolytica ?
-
 ?
Laminin-1 + H2O
-
 Entamoeba histolytica HM1-IMSS ?
-
 ?
additional information recombinant CP4 does not cleave benzyloxycarbonyl-RR-7-amido-4-methylcoumarin or benzyloxycarbonyl-FR-7-amido-4-methylcoumarin Entamoeba histolytica ?
-
 ?
additional information recombinant CP4 does not cleave benzyloxycarbonyl-RR-7-amido-4-methylcoumarin or benzyloxycarbonyl-FR-7-amido-4-methylcoumarin Entamoeba histolytica HM1-IMSS ?
-
 ?
pro-interleukin-18 + H2O
-
 Entamoeba histolytica ?
-
 ?
Protein + H2O CP4 displays a unique preference for valine and isoleucine at P2 (Val > Ile) and broader specificities at the P1 and P3 positions, while at P4 proline is relatively selective Entamoeba histolytica ?
-
 ?
Protein + H2O CP4 displays a unique preference for valine and isoleucine at P2 (Val > Ile) and broader specificities at the P1 and P3 positions, while at P4 proline is relatively selective Entamoeba histolytica HM1-IMSS ?
-
 ?
succinyl-LLVY-7-amido-4-methylcoumarin + H2O
-
 Entamoeba histolytica succinyl-LLVY + 7-amino-4-methylcoumarin
-
 ?
succinyl-LLVY-7-amido-4-methylcoumarin + H2O
-
 Entamoeba histolytica HM1-IMSS succinyl-LLVY + 7-amino-4-methylcoumarin
-
 ?
villin-1 + H2O
-
 Entamoeba histolytica ?
-
 ?

Synonyms

Synonyms Comment Organism
CP4
-
 Entamoeba histolytica
cysteine proteinase 4
-
 Entamoeba histolytica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 the optimum for substrate cleavage is at pH 7.0 Entamoeba histolytica

pH Stability

pH Stability pH Stability Maximum Comment Organism
3 4 recombinant CP4 is efficiently autoactivated at pH 3.0-4.0 with 5 mM dithiothreitol, mature recombinant CP4 is unstable at acid pH and undergoes further degradation Entamoeba histolytica

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.00967
-
 WRR605 apparent value, pH 7.0, at 37°C Entamoeba histolytica

Expression

Organism Comment Expression
Entamoeba histolytica co-culture of Entamoeba histolytica trophozoites with mucin-producing T84 cells increases ehcp4 expression up to 6fold up