Information on EC 3.4.22.B14 - papain-like proteinase 1

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.B14
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
papain-like proteinase 1
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
papain-like proteinase 1 is responsible for cleavages located at the N-terminus of the replicase polyprotein
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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-
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain Ent
SwissProt
Manually annotated by BRENDA team
strain Ent
SwissProt
Manually annotated by BRENDA team
strains 229E, NL, OC43 and HKU1
SwissProt
Manually annotated by BRENDA team
strain Beaudette
SwissProt
Manually annotated by BRENDA team
strain Beaudette
SwissProt
Manually annotated by BRENDA team
strain A59
SwissProt
Manually annotated by BRENDA team
strain JHM
SwissProt
Manually annotated by BRENDA team
strain A59
SwissProt
Manually annotated by BRENDA team
severe acute respiratory syndrome coronavirus, Urbani strain
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-
Manually annotated by BRENDA team
strain Purdue 46-MAD
SwissProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-[[4-(dimethylamino)phenyl]azo]benzoic acid-MYNKMGGGDKTVSF-(5-[(2-aminoethyl)amino]naphthalene-1-sulfonic acid) amide + H2O
?
show the reaction diagram
coronavirus nonstructural protein 1 + H2O
?
show the reaction diagram
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-
-
-
?
coronavirus nonstructural protein 2 + H2O
?
show the reaction diagram
-
-
-
-
?
coronavirus nonstructural protein 3 + H2O
?
show the reaction diagram
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-
-
-
?
L-Arg-L-Leu-L-Arg-Gly-Gly-7-amido-4-methylcoumarin + H2O
L-Arg-L-Leu-L-Arg-Gly-Gly + 7-amino-4-methylcoumarin
show the reaction diagram
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-
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-
?
ORF 1a polyprotein + H2O
?
show the reaction diagram
ORF 1a polyprotein + H2O
nsp1 + nsp2 + nsp3
show the reaction diagram
p28
?
show the reaction diagram
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-
-
-
?
p65
?
show the reaction diagram
-
-
-
-
?
ubiquitin-7-amido-4-methylcoumarin + H2O
ubiquitin + 7-amino-4-methylcoumarin
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ORF 1a polyprotein + H2O
?
show the reaction diagram
p28
?
show the reaction diagram
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-
-
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?
p65
?
show the reaction diagram
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-
-
-
?
additional information
?
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,6-dimethyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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2,N-dimethyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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2-methoxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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2-methoxymethyl-5-nitrobenzoic acid methyl ester
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2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
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2-methyl-N-[1-(naphthalen-2-yl)ethyl]benzamide
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3-methoxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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3-methyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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4-amino-N-[(R)-1-(1-naphthyl)ethyl]benzamide
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4-amino-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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4-methoxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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4-methyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
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5-amino-2-methoxymethyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
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5-amino-2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
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5-amino-2-methyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
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5-cyano-2-methyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
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5-iodo-2-methyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
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5-methylamino-2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
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5-N-acetylamino-2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
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5-N-tert-butoxycarbonylmethylamino-2-methyl-N'-[(R)-1-(1-naphthyl)ethyl]benzamide
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E-64d
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membrane-permeable do the irreversible cysteine proteinase inhibitor E-64 blocks cleavage of p65 in vivo
leupeptin
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blocks the ability of PLP1 to cleave p28 in vitro but not in vivo due to permeability and toxicity issues
N-methyl-5-methylamino-2-methyl-N'-[(R)-1-(1-naphthyl)ethyl]benzamide
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ZnCl2
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blocks the ability of PLP1 to cleave p28 in vitro but not in vivo due to permeability and toxicity issues
additional information
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not inhibited by 2-hydroxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide, 4-N'-tert-butoxycarbonylamino-N-[(R)-1-(2-naphthyl)ethyl]-benzamide, 2-methyl-N-[1-(2-naphthyl)propyl]benzamide, and 2-methyl-N-[1-(2-naphthyl)benzyl]benzamide
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0121
2,6-dimethyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0226
2,N-dimethyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.09
2-methoxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0014
2-methoxymethyl-5-nitrobenzoic acid methyl ester
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0023
2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0087
2-methyl-N-[1-(naphthalen-2-yl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0135
3-methoxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0148
3-methyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0248
4-amino-N-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0461
4-amino-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.149
4-methoxy-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0291
4-methyl-N-[(R)-1-(2-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0052
5-amino-2-methoxymethyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.00056
5-amino-2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.00056
5-amino-2-methyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0111
5-cyano-2-methyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0027
5-iodo-2-methyl-N-[1-methyl-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.00046
5-methylamino-2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.00264
5-N-acetylamino-2-methyl-N-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0048
5-N-tert-butoxycarbonylmethylamino-2-methyl-N'-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
0.0013
N-methyl-5-methylamino-2-methyl-N'-[(R)-1-(1-naphthyl)ethyl]benzamide
SARS coronavirus
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in 50 mM HEPES, pH 7.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
reaction is significantly more efficient at 22C than when carried out at 30C
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 0.1-0.18 M (NH4)2SO4, 0.1 M MES (pH 6.5), and 30% (w/v) PEG 5000, at 19C
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography and Superdex 75 gel filtration
recombinant enzyme
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned into plasmids under T7 promoter control and cotransfected into HeLa-MHVR cells infected with recombinant vaccinia virus
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expressed in Escherichia coli
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expressed in Escherichia coli Rosetta (DE3)pLysS cells
expression in Escherichia coli by using a T7 RNA polymerase promoter system or as a maltose-binding protein fusion protein. With both overexpression systems the recombinant PLP-1 exhibits trans cleavage activity. When PLP-1 is expressed as a polypeptide that includes additional viral sequences at the carboxyl terminus of the predicted PLP-1 domain, a fivefold increase in proteolytic activity is observed