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Information on EC 3.4.22.B78 - endopepidase PH1704 and Organism(s) Pyrococcus horikoshii and UniProt Accession O59413

for references in articles please use BRENDA:EC3.4.22.B78
preliminary BRENDA-supplied EC number
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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.22 Cysteine endopeptidases
                3.4.22.B78 endopepidase PH1704
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This record set is specific for:
Pyrococcus horikoshii
UNIPROT: O59413
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The taxonomic range for the selected organisms is: Pyrococcus horikoshii
The expected taxonomic range for this enzyme is: Pyrococcus horikoshii
Reaction Schemes
hide
the enzyme prefers substrates with an Arg residue at the P1 site
Synonyms
PH1704, PH1704 protease, PhpI, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
PH1704 protease
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
the enzyme prefers substrates with an Arg residue at the P1 site
show the reaction diagram
it also acts as arginyl aminopeptidase with higher efficiency
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Ala-Ala-Leu-Arg-7-amido-4-methylcoumarin + H2O
Ala-Ala-Leu-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Ala-Ala-Phe-Arg-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
Ala-Arg-7-amido-4-methylcoumarin + H2O
Ala-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
Leu-Arg-7-amido-4-methylcoumarin + H2O
Leu-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
additional information
?
-
the enzyme has both aminopeptidase and endopeptidase activities, substrate specificity, overview. Ala-Phe-7-amido-4-methylcoumarin, and Ala-Ala-Phe-7-amido-4-methylcoumarin are poor substrates. Phe-7-amido-4-methylcoumarin, Ala-7-amido-4-methylcoumarin, Val-7-amido-4-methylcoumarin, Asp-7-amido-4-methylcoumarin, and Ser-7-amido-4-methylcoumarin are no substrates
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
5 mM, 29% inhibition of protease activity
Co2+
0.2 mM, 75% inhibition of protease activity
Cu2+
0.2 mM, 57% inhibition of protease activity
Fe3+
0.2 mM, 27% inhibition of protease activity
K+
5 mM, 19% inhibition of protease activity
Mg2+
5 mM, 10% inhibition of protease activity
Mn2+
5 mM, 81% inhibition of protease activity
Na+
5 mM, 28% inhibition of protease activity
Ni2+
0.2 mM, 84% inhibition of protease activity
Zn2+
0.2 mM, 63% inhibition of protease activity
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
inhibits 29% at 5 mM, 19% at 1 mM
Co2+
inhibits 74% at 0.2 mM and precipitates the enzyme at 1 mM
Cu2+
inhibits 57% at 0.2 mM, 81% at 1 mM, and precipitates the enzyme at 5 mM
Fe3+
inhibits 28% at 0.2 mM, 68% at 1 mM, and precipitates the enzyme at 5 mM
iodoacetamide
K+
inhibits 19% at 5 mM
Mg2+
inhibits 10% at 5 mM
Mn2+
inhibits 81% at 5 mM, 42% at 1 mM
Na+
inhibits 28% at 5 mM
Ni2+
inhibits 85% at 0.2 mM and precipitates the enzyme at 1 mM
Zn2+
inhibits 64% at 0.2 mM, 77% at 1 mM, and precipitates the enzyme at 5 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
steady-state kinetics, recombinant wild-tyype and mutant enzymes, overview
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
19.87
purified recombinant wild-type enzyme, pH 7.5, 85°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
pH 6.5: about 45% of maximal activity, pH 9.0: about 55% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50 - 95
50°C: about 60% of maximal activity, 95°C: about 60% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme is a member of DJ-1/ThiJ/PfpI superfamily that has diverse functional subclasses
additional information
the enzyme has both aminopeptidase and endopeptidase activities, Cys100 is the catalytic nucleophilic residue, residue Tyr120 is important in substrate binding and is involved in enzyme activity to form a hydrogen bond with Cys100 and as an entrance gate of the substrate with Lys43, active site pocket structure, molecular docking study, overview. Cys100, His101, and Glu474 function as a catalytic triad. Active residues are Glu12, Glu15, Lys43, Gly70, Arg71, Cys100, His101, Tyr120, Val150, Arg471, Glu474, and Arg475
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000
12 * 20000, about, recombinant enzyme, SDS-PAGE
240000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dodecamer
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E12T
the mutant enzyme is more stable than the wild-type enzyme
Y120P
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by 8fold heat treatment at 80°C for 15 min, and gel filtration. Affinity chromatography is not suitable for this enzyme because recombinant PH1704 can be precipitated in the present of nickel
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, phylogenetic tree, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Zhan, D.; Han, W.; Feng, Y.
Experimental and computational studies indicate the mutation of Glu12 to increase the thermostability of oligomeric protease from Pyrococcus horikoshii
J. Mol. Model.
17
1241-1249
2011
Pyrococcus horikoshii (O59413), Pyrococcus horikoshii DSM 12428 (O59413)
Manually annotated by BRENDA team
Zhan, D.; Bai, A.; Yu, L.; Han, W.; Feng, Y.
Characterization of the PH1704 protease from Pyrococcus horikoshii OT3 and the critical functions of Tyr120
PLoS One
9
e103902
2014
Pyrococcus horikoshii (O59413), Pyrococcus horikoshii DSM 12428 (O59413), Pyrococcus horikoshii OT-3 (O59413)
Manually annotated by BRENDA team