Information on EC 3.4.22.B50 - papain-like proteinase 2

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.B50
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
papain-like proteinase 2
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
responsible for the cleavages located at the N-terminus of the replicase polyprotein
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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-
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CAS REGISTRY NUMBER
COMMENTARY hide
851883-30-2
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SwissProt
Manually annotated by BRENDA team
i.e. mouse hepatitis virus
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E-EDANS)RELNGGAPI(K-DABCYL)S + H2O
?
show the reaction diagram
Abz-FRLKGGAPIKGV-N-(2,4-dinitrophenyl)-ethylenediamine + H2O
?
show the reaction diagram
azocasein + H2O
?
show the reaction diagram
-
-
-
?
BCoV substrate
?
show the reaction diagram
branched polyubiquitin chains + H2O
?
show the reaction diagram
diubiquitin + H2O
?
show the reaction diagram
FRLKGG-4-nitroanilide + H2O
FRLKGG + 4-nitroaniline
show the reaction diagram
-
-
-
?
FRLKGGAPIKGV
?
show the reaction diagram
FTKLAGGKISFS + H2O
FTKLAG + GKISFS
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
ISG15-nsp2-fusion protein + H2O
?
show the reaction diagram
ISLKGGKIVSTC
?
show the reaction diagram
MHV substrate
?
show the reaction diagram
-
-
-
-
?
N-benzoyl-Phe-Val-Arg-4-nitroanilide + H2O
N-benzoyl-Phe-Val-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
?
PLP2-MP1 precursor polyprotein + H2O
MP1 + ?
show the reaction diagram
-
-
-
-
?
polyubiquitin + H2O
monoubiquitin + ?
show the reaction diagram
the enzyme cleaves Lys48- and Lys63-linked polyubiquitin to monoubiquitin but not linear polyubiquitin
-
-
?
RELNGGAVTRYV + H2O
AVTYRV + RELNGG
show the reaction diagram
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12 mer oligopeptide containing Gly180-Ala181, 5% cleavage
-
-
?
replicase polyprotein + H2O
?
show the reaction diagram
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PLP2 cleaves a substrate encoding the first predicted membrane-spanning domain, MP1, of the replicase polyprotein. Processing the replicase polyprotein at this site generates the p150 replicase intermediate that is likely critical for embedding the replicase complex into cellular membranes. The enzyme acts efficiently in trans
-
-
?
RLRGG-7-amido-4-methylcoumarin + H2O
RLRGG + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
ubiquitin + H2O
?
show the reaction diagram
ubiquitin-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
ubiquitin-7-amido-4-trifluoromethylcoumarin + H2O
ubiquitin + 7-amino-4-trifluoromethylcoumarin
show the reaction diagram
the enzyme catalyzes proteolytic processing of the viral polyprotein and also shows significant in vitro deubiquitinating and de-ISGylating activities. The enzyme binds ubiquitin, the ubiquitin core makes mostly hydrophilic interactions with the enzyme, while the Leu-Arg-Gly-Gly C-terminus of ubiquitin is located in the catalytic cleft of the enzyme. The ubiquitin core binds to the palm, thumb and fingers domains of the enzyme, while its final four C-terminal residues bind into a narrow channel by a network of hydrogen bonds and reach towards the active site
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-
?
ubiquitin-aminomethylcoumarin + H2O
?
show the reaction diagram
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-
-
?
ubiquitinated branched peptides + H2O
?
show the reaction diagram
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-
-
-
?
ubiquitinated RIG-I + H2O
deubiquitinated RIG-I + ubiquitin
show the reaction diagram
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-
-
?
ubiquitinated STING + H2O
deubiquitinated STING + ubiquitin
show the reaction diagram
-
-
-
?
VAKQGAGFKRTY + H2O
VAKQGA + GFKRTY
show the reaction diagram
-
-
-
-
?
viral replicase polyprotein + H2O
?
show the reaction diagram
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PLP2 is responsible for processing both cleavage sites 2 and 3 to release nsp2 and nsp3. The cleavage sites are identified as FTKLAG-/-GKISFS for CS2 and VAKQGA-/-GFKRTY for CS3.
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-
?
Z-KKAG-7-amido-4-methylcoumarin + H2O
Z-KKAG + 7-amino-4-methylcoumarin
show the reaction diagram
the catalytic efficiency toward Z-KKAG-7-amido-4-methylcoumarin is 5times higher than that for Z-LRGG-7-amido-4-methylcoumarin
-
-
?
Z-LRGG-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
Z-LRGG-7-amido-4-methylcoumarin + H2O
Z-LRGG + 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
FTKLAGGKISFS + H2O
FTKLAG + GKISFS
show the reaction diagram
-
-
-
-
?
polyubiquitin + H2O
monoubiquitin + ?
show the reaction diagram
P0C6Y1
the enzyme cleaves Lys48- and Lys63-linked polyubiquitin to monoubiquitin but not linear polyubiquitin
-
-
?
replicase polyprotein + H2O
?
show the reaction diagram
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PLP2 cleaves a substrate encoding the first predicted membrane-spanning domain, MP1, of the replicase polyprotein. Processing the replicase polyprotein at this site generates the p150 replicase intermediate that is likely critical for embedding the replicase complex into cellular membranes. The enzyme acts efficiently in trans
-
-
?
ubiquitin + H2O
?
show the reaction diagram
ubiquitinated RIG-I + H2O
deubiquitinated RIG-I + ubiquitin
show the reaction diagram
P0C6Y4
-
-
-
?
ubiquitinated STING + H2O
deubiquitinated STING + ubiquitin
show the reaction diagram
P0C6Y4
-
-
-
?
VAKQGAGFKRTY + H2O
VAKQGA + GFKRTY
show the reaction diagram
-
-
-
-
?
additional information
?
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the enzyme is one of three distinct viral proteases (PLP1, PLP2 and 3CLpro) involved in processing of the replicase polyprotein
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
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the fingers domain of the enzyme contains a zinc ion that is tetrahedrally coordinated by four cysteines
additional information
-
no effect by Mg2+, Ca2+, Mn2+, Co2+, and Ni2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(4E)-1,7-bis(3,4-dihydroxyphenyl)hept-4-en-3-one
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a natural diarylheptanoide inhibitor
(7R)-5,7-dihydroxy-8-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-methylpentyl)-2-methyl-3,4,7,8-tetrahydro-2H,6H-pyrano[3,2-g]chromen-6-one
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a natural geranylated flavonoid inhibitor
1,6,6-trimethyl-1,2,6,7,8,9-hexahydrophenanthro[1,2-b]furan-10,11-dione
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a natural tanshinone inhibitor
2-methyl-N-[(1R)-1-(naphthalen-2-yl)ethyl]benzamide
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over 90% inhibition at 0.1 mM
2-methyl-N-[(1S)-1-(naphthalen-2-yl)ethyl]benzamide
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14% inhibition at 0.1 mM
2-methyl-N-[1-(naphthalen-2-yl)ethyl]benzamide
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racemat
5-amino-2-methyl-N-[(1R)-1-(naphthalen-1-yl)ethyl]benzamide
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R-isomer, a potent, noncovalent, competitive inhibitor
6-Mercaptopurine
6-Thioguanine
hydroxypridine-2-thione Zn
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hydroxypyridine-2-thione-Zn(II)
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N-(1,3-benzodioxol-5-ylmethyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
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N-(3-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
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N-(4-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
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N-(4-methoxybenzyl)-1-(naphthalen-1-ylmethyl)piperidine-4-carboxamide
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N-cyclohexyl-2-aminethanesulfonic acid
the molecule binds near the catalytic triad
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N-ethyl-N-phenyldithiocarbaic acid Zn
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N-ethyl-N-phenyldithiocarbamic acid-Zn(II)
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N-[(2-methoxypyridin-4-yl)methyl]-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
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N-[3-(acetylamino)benzyl]-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
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NSC158011
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Ub-3Br
suicide inhibitor, i.e. ubiquitin(1-75)-3-bromopropylamide
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Ubiquitin aldehyde
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-
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additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
L-cysteine
20 mM used in assay conditions
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.021 - 0.0772
Abz-FRLKGGAPIKGV-N-(2,4-dinitrophenyl)-ethylenediamine
0.185
FRLKGGAPIKGV
-
-
0.09687 - 0.1769
N-benzoyl-Phe-Val-Arg-4-nitroanilide
0.73 - 3.33
ubiquitin-7-amido-4-trifluoromethylcoumarin
-
3.23
Z-KKAG-7-amido-4-methylcoumarin
in 20 mM Tris-HCl, pH 7.5, 0.1 mg/ml BSA, 150 mM NaCl, 2 mM dithiothreitol, at 25°C
0.67
Z-LRGG-7-amido-4-methylcoumarin
in 20 mM Tris-HCl, pH 7.5, 0.1 mg/ml BSA, 150 mM NaCl, 2 mM dithiothreitol, at 25°C
additional information
additional information
Michaelis-Menten steady-state enzyme-kinetics of wild-type and mutant enzymes, overview
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.023 - 0.366
Abz-FRLKGGAPIKGV-N-(2,4-dinitrophenyl)-ethylenediamine
0.025
FRLKGGAPIKGV
Alphacoronavirus
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-
0.23 - 2.65
N-benzoyl-Phe-Val-Arg-4-nitroanilide
0.01 - 0.79
ubiquitin-7-amido-4-trifluoromethylcoumarin
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.8 - 14.87
N-benzoyl-Phe-Val-Arg-4-nitroanilide
31569
0.045 - 0.155
RLRGG-7-amido-4-methylcoumarin
202546
0.01 - 1.08
ubiquitin-7-amido-4-trifluoromethylcoumarin
202465
0.413 - 17
ubiquitin-aminomethylcoumarin
202547
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00049
5-amino-2-methyl-N-[(1R)-1-(naphthalen-1-yl)ethyl]benzamide
-
pH and temperature not specified in the publication
0.015
6-Mercaptopurine
pH 7.4, 30°C
0.015
6-Thioguanine
pH 7.4, 30°C
0.00021
Ubiquitin aldehyde
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-
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IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0041
(4E)-1,7-bis(3,4-dihydroxyphenyl)hept-4-en-3-one
SARS coronavirus
-
pH and temperature not specified in the publication
0.005
(7R)-5,7-dihydroxy-8-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-4-methylpentyl)-2-methyl-3,4,7,8-tetrahydro-2H,6H-pyrano[3,2-g]chromen-6-one
SARS coronavirus
-
pH and temperature not specified in the publication
0.0008
1,6,6-trimethyl-1,2,6,7,8,9-hexahydrophenanthro[1,2-b]furan-10,11-dione
SARS coronavirus
-
pH and temperature not specified in the publication
0.0087
2-methyl-N-[(1R)-1-(naphthalen-2-yl)ethyl]benzamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.0201
2-methyl-N-[1-(naphthalen-2-yl)ethyl]benzamide
SARS coronavirus
-
racemate, pH and temperature not specified in the publication
0.0006
5-amino-2-methyl-N-[(1R)-1-(naphthalen-1-yl)ethyl]benzamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.0216
6-Mercaptopurine
SARS coronavirus
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pH and temperature not specified in the publication
0.005
6-Thioguanine
SARS coronavirus
-
pH and temperature not specified in the publication
0.0001712 - 0.000349
E-64
0.0037
hydroxypyridine-2-thione-Zn(II)
SARS coronavirus
-
pH and temperature not specified in the publication
-
0.00032
N-(1,3-benzodioxol-5-ylmethyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.00015
N-(3-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.00049
N-(4-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.059
N-(4-methoxybenzyl)-1-(naphthalen-1-ylmethyl)piperidine-4-carboxamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.0033
N-ethyl-N-phenyldithiocarbamic acid-Zn(II)
SARS coronavirus
-
pH and temperature not specified in the publication
-
0.00035
N-[(2-methoxypyridin-4-yl)methyl]-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
SARS coronavirus
-
pH and temperature not specified in the publication
0.00039
N-[3-(acetylamino)benzyl]-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide
SARS coronavirus
-
pH and temperature not specified in the publication
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30.78
wild type enzyme, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C
63.11
mutant enzyme S32T/A67Y, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C
73.63
mutant enzyme S32T, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C
86.95
mutant enzyme A67Y, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 8.6
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-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 8
more than 50% activity between pH 4.5 and 8.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
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SDS-PAGE and Western blot detection with His6-specific antibodies, P-Plpro(C1)-His6 fragment
40000
x * 40000, SDS-PAGE
41000
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SDS-PAGE and Western blot detection with His6-specific antibodies, P-Plpro(C2)-His6 fragment
51000
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SDS-PAGE
56700
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gel filtration
60000
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SDS-PAGE and Western blot detection with His6-specific antibodies, T7-ISG15P-Plpro(C2)-His6 fragment
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 40000, SDS-PAGE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapor-diffusion, to 1.85 A resolution, the enzyme has an intact zinc-binding motif, an unobstructed catalytically competent active site and an intriguing, ubiquitin-like N-terminal domain
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in complex with ubiquitin, hanging drop vapor diffusion method, using 100 mM MES, pH 6.2, 18% (w/v) polyethylene glycol 20,000
hanging drop vapor diffusion method, using 100 mM Bis-Tris, pH 6.5, 25-30% (w/v) polyethylene glycol monomethyl ether 2000
apoenzyme and enzyme in complexes with ubiquitin, and inhibitors 5-amino-2-methyl-N-[(1R)-1-(naphthalen-1-yl)ethyl]benzamide, N-(1,3-benzodioxol-5-ylmethyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide, N-(4-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide, and N-(3-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide, sitting drop vapor diffusion method, apoenzyme by mixing of 1.1-20 mg/ml protein in 20 mM Tris, pH 7.5, 10 mM DTT, with 100 mM sodium citrate, pH 5.2, 3 M ammonium sulfate. Enzyme-inhibitor N-(1,3-benzodioxol-5-ylmethyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide complex by mixing of 5 mg/ml protein in 20 mM Tris, pH 7.5, 10 mM DTT, with 1 mM inhibitor, 1 M (NH4)2SO4, 50 mM MES, pH 6.5, and 2.5% PEG 400. Enzyme-inhibitor 5-amino-2-methyl-N-[(1R)-1-(naphthalen-1-yl)ethyl]benzamide by mixing of 8 mg/ml protein in 20 mM Tris, pH 7.5, 10 mM DTT with 0.20 mM inhibitor, 1 M LiCl2, 0.1 M MES pH 6.0, 30% PEG 6000. Enzyme-inhibitor N-(3-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide or N-(4-fluorobenzyl)-1-[(1R)-1-(naphthalen-1-yl)ethyl]piperidine-4-carboxamide complex by mixing of 6 and 12 mg/ml protein, respectively, in 25 mM Tris, pH 7.5, 100 mM NaCl, 10 mM DTT with 0.20 mM inhibitor, 100 mM sodium citrate, pH 5.5, 40% v/v PEG 600. Enzyme-ubiquitin complex by mixing of 3-12 mg/ml protein and ubiquitin in 20 mM Tris, pH 7.5, with 0.1 M CHES, pH 9.5, 18% PEG 3000, X-ray diffraction structure determination and analysis at 1.4-2.75 A resolution
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purified enzyme mutant C112S in complex with ubiquitin, sitting drop vapour diffusion method, mixing of enzyme and ubiquitin in a 1:1 molar ratio giving 8 mg/ml and 2 mg/ml, respectively, addition of a reservoir solution consisting of 18% PEG 3000, 0.1 M CHES, pH 9.5, 22°C, X-ray diffraction structure determination and analysis at 1.4 A resolution
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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thermolabil, significantly reduced activity after incubation at temperatures higher than 37°C for 30 min
40 - 80
after 10 incubation, the enzyme shows 100% activity between 40 and 55°C, about 97% activity at 60°C, about 65% activity at 70°C, 50% activity at 72°C, about 25% activity at 75°C, and no activity at 80°C
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, Tris buffer, pH 7.5, 20% glycerol
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-80°C, Tris-HCl buffer, pH 8.0, 10% glycerol, 1 month
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4°C, Tris-HCl buffer, pH 8.0, 10% glycerol, 24 h
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by His-Bind Purification Kit
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gel filtration
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GST resin column chromatography, Mono Q column chromatography, and Superdex 75 gel filtration
Ni-NTA column chromatography
Ni-NTA column chromatography, Superdex 75 gel filtration, and Source 15Q column chromatography
purified to homogeneity by hydrophobic interaction and anion exchange chromatography
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recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3), the secretion tag is removed and the purification continued by nickel affinity chromatography, gel filtration, and ultrafiltration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned into modified version of the pBacPAK8/His2 vector and expressed in insect cells Sf9
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21-Gold (DE3) cells
expressed in HEK-293T cells
expression in Escherichia coli BL21 (DE3) and HeLa cells
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expression in Escherichia coli XL-1 Blue and HeLa-MHV receptor cells
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recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes, with a secretion tag, in Escherichia coli strain BL21(DE3)
vaccinia-T1-mediated expression of a cDNA clone encoding the MHV PLP2 domain
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C1651A
C1732A
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encodes a substitution of a cysteine residue, predicted to be critical for zinc binding
D1826A
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purified SARS-CoV PLpro protein containing an alanine substitution at putative catalytic residues
C1651A
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Plpro active-site mutant; purified SARS-CoV PLpro protein containing an alanine substitution at putative catalytic residues
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C1732A
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encodes a substitution of a cysteine residue, predicted to be critical for zinc binding
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D1826A
-
purified SARS-CoV PLpro protein containing an alanine substitution at putative catalytic residues
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C270A/H332A
active site mutant
I353R
the mutant exhibits about 40fold reduction in specificity toward ubiquitin compared to the wild type while the activity toward RLRGG-7-amido-4-methylcoumarin is essentially unaltered
I353W
the mutant exhibits about 10fold reduction in specificity toward ubiquitin compared to the wild type while the activity toward RLRGG-7-amido-4-methylcoumarin is essentially unaltered
T312A/I313V/I353R
the mutant shows the greatest decrease in inhibitory activity in the interferon-beta promoter activity assay
C106A
-
inactive
C1729A
the mutant shows reduced interferon antagonistic activity compared to that of the wild type enzyme
D1901A
the mutant almost completely loses interferon antagonistic activity compared to that of the wild type enzyme
H1888A
the mutant almost completely loses interferon antagonistic activity compared to that of the wild type enzyme
D165A
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
E168A
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
E168D
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
E168R
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
L163Q
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
Y265A
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
Y265F
site-directed mutagenesis, the mutant shows reduced activity in deubiquitination compared to the wild-type enzyme
Y274A
site-directed mutagenesis, inactive mutant
A67Y
the mutation enhances the catalytic efficiency of the enzyme about 8fold while the thermostability of the mutant enzyme remains unchanged. The specific activity against azo-casein is almost 2.4 times higher than wild type
S32T
the mutation enhances the catalytic efficiency of the enzyme about 8fold while the thermostability of the mutant enzyme remains unchanged
S32T/A67Y
the mutations enhance the catalytic efficiency of the enzyme about 8fold while the thermostability of the mutant enzyme remains unchanged
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
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the enzyme is an attractive antiviral drug target because it is essential for coronaviral replication. Targeting the enzyme with antiviral drugs may have an advantage in not only inhibiting viral replication but also inhibiting the dysregulation of signaling cascades in infected cells that may lead to cell death in surrounding, uninfected cells
medicine