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 | 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 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 | 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
23000 | - |
calculated molecular mass of the mature enzyme | Entamoeba histolytica |
26000 | - |
active recombinant CP4, SDS-PAGE | Entamoeba histolytica |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Entamoeba histolytica | C4LTT0 | - |
- |
Entamoeba histolytica HM1-IMSS | C4LTT0 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel affinity column chromatography | Entamoeba histolytica |
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 | Comment | Organism | Textmining |
---|---|---|---|
trophozoite | - |
Entamoeba histolytica | - |
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 | Comment | Organism |
---|---|---|
CP4 | - |
Entamoeba histolytica |
cysteine proteinase 4 | - |
Entamoeba histolytica |
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 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 maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00967 | - |
WRR605 | apparent value, pH 7.0, at 37°C | Entamoeba histolytica |
Organism | Comment | Expression |
---|---|---|
Entamoeba histolytica | co-culture of Entamoeba histolytica trophozoites with mucin-producing T84 cells increases ehcp4 expression up to 6fold | up |