Information on EC 3.4.22.29 - picornain 2A

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.29
-
RECOMMENDED NAME
GeneOntology No.
picornain 2A
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
selective cleavage of Tyr-/-Gly bond in picornavirus polyprotein
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
103406-62-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
Coxsackievirus B3 Woodruff
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
hepatitis A virus
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
human rhinovirus C2 W12
-
UniProt
Manually annotated by BRENDA team
poliovirus 2A
-
-
-
Manually annotated by BRENDA team
strain J1
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
a PV 2Apro variant deficient in eukaryotic initiation factor (eIF) 4GI cleavage does not increase picornavirus IRES-driven translation
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1C'D2A precursor polypeptide + H2O
?
show the reaction diagram
-
i.e. autocatalytic cis cleavage activity, cleaves at 1D/2A junction in very rapid cotranslational reaction
-
-
?
2-aminobenzoic acid-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Phe(NO2)-Ala-OH + H2O
?
show the reaction diagram
-
-
-
-
?
3CD-precursor poliovirus protein + H2O
poliovirus 3C' and 3D'-protein
show the reaction diagram
-
-
-
?
acetyl-LSTT-7-amido-4-trifluoromethylcoumarin + H2O
acetyl-LSTT + 7-amino-4-trifluoromethylcoumarin
show the reaction diagram
-
-
-
-
?
Arg-Arg-Asn-Thr-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Arg-Arg-Asn-Thr-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
peptide derived from poliovirus type 1 polyprotein, poor substrate
-
?
Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
i.e. synthetic peptide P7-P8', hydrolyzed with about the same relative efficiency compared to P8-P8'
-
?
Bid protein + H2O
?
show the reaction diagram
-
-
-
-
?
cAMP-regulated response element binding protein + H2O
?
show the reaction diagram
-
-
-
-
?
capsid precursor protein P1 + H2O
viral protein 1ABC + viral protein 1D
show the reaction diagram
-
-
-
?
cellular eukaryotic translation initiation factor eIF4GI + H2O
peptides
show the reaction diagram
-
-
-
-
?
cellular eukaryotic translation initiation factor eIF4GII + H2O
peptides
show the reaction diagram
-
-
-
-
?
CVB1 protein + H2O
?
show the reaction diagram
-
-
-
-
?
cytokeratin 8 + H2O
?
show the reaction diagram
DIKSYGLGPRYGG + H2O
DIKSY + GLGPRYGG
show the reaction diagram
-
-
-
-
?
dystrophin + H2O
?
show the reaction diagram
eIF4G + H2O
?
show the reaction diagram
eIF4G + H2O
fragments of eIF4G
show the reaction diagram
-
-
-
-
?
eIF4GI + H2O
fragments of eIF4GI
show the reaction diagram
-
-
-
-
?
eukaryotic initiation factor 4G + H2O
?
show the reaction diagram
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
eukaryotic translation initiation factor 4G I + H2O
?
show the reaction diagram
-
-
-
?
eukaryotic translation initiation factor 4G II + H2O
?
show the reaction diagram
-
-
-
?
eukaryotic translation initiation factor 4gamma + H2O
?
show the reaction diagram
-
-
-
-
?
eukaryotic translation initiation factor 4Gl + H2O
?
show the reaction diagram
-
-
-
-
?
Glu-Arg-Ala-Ser-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Glu-Arg-Ala-Ser-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
peptide derived from poliovirus type 1 polyprotein
-
?
Gly-Leu-Gly-Gln-Met methyl ester + H2O
Gly-Leu-Gly-Gln-Met + CH3OH
show the reaction diagram
-
esterase activity
-
?
GRTTLST-(3-nitrotyrosine)-GPPR-(lysine anthranilide)-Y + H2O
GRTTLST-(3-nitrotyrosine) + GPPR-(lysine anthranilide)-Y
show the reaction diagram
-
-
-
-
?
Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
i.e. synthetic peptide P5-P8', poor substrate
-
?
Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
i.e. synthetic peptide P4-P8', poor substrate
-
?
KSYKVSTSGPRAFSSR + H2O
KSYKVSTS + GPRAFSSR
show the reaction diagram
L-Leu-Val-Pro-Arg-Gly-Ser + H2O
L-Leu-Val-Pro-Arg + Gly-Ser
show the reaction diagram
-
-
-
-
?
modified pentadecameric peptides + H2O
?
show the reaction diagram
-
i.e. synthetic P8-P7' peptides, intermolecular specificity, changes at P2 and P1' are highly deleterious
-
-
?
mouse double minute 2 + H2O
?
show the reaction diagram
-
-
-
-
?
mouse double minute 4 + H2O
?
show the reaction diagram
-
-
-
-
?
oligopeptides corresponding to cleavage sites of coxsackievirus + H2O
?
show the reaction diagram
-
-
-
-
?
oligopeptides corresponding to cleavage sites of human rhinovirus + H2O
?
show the reaction diagram
-
-
-
-
?
oligopeptides corresponding to cleavage sites of poliovirus type 1 + H2O
?
show the reaction diagram
oligopeptides derived from eIF-4gamma + H2O
?
show the reaction diagram
-
human rhinovirus, common cleavage site: Ala-Gly
-
-
?
P1/P2 precursor polypeptide + H2O
poliovirus 1C'D polypeptide + proteinase 2Apro
show the reaction diagram
picornavirus polyprotein + H2O
?
show the reaction diagram
-
intramolecular reaction, processing begins before synthesis of the polyprotein is complete, cleavage separates capsid protein precursor and noncapsid protein precursor
-
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
poliovirus polypeptide + H2O
?
show the reaction diagram
poliovirus polyprotein fragment + H2O
hydrolyzed poliovirus polyprotein fragment
show the reaction diagram
-
containing cleavage site at P1/P2 junction
partially cleaved in trans
?
poly(A) binding protein + H2O
fragments of poly(A) binding protein
show the reaction diagram
-
contains 1 cleavage site for 2A proteinase within the proline-rich linker domain
-
-
?
poly-(ADP-ribose) polymerase + H2O
?
show the reaction diagram
-
-
-
-
?
Pro-Arg-Ala-Ser-Met-Lys-Thr-Val-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Pro-Arg-Ala-Ser-Met-Lys-Thr-Val + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
poor substrate
-
?
Pro-Arg-Glu-Asn-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Pro-Arg-Glu-Asn-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
peptide derived from poliovirus type 1 polyprotein, poor substrate
-
?
pro-caspase 3 + H2O
caspase 3 + ?
show the reaction diagram
-
-
-
-
?
Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met + H2O
Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met
show the reaction diagram
-
i.e. synthetic peptide P6-P5', hydrolyzed with 42% relative efficiency compared to P8-P8'
-
?
Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr + H2O
Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr
show the reaction diagram
-
i.e. synthetic peptide P6-P6', hydrolyzed with 45% relative efficiency compared to P8-P8', smallest cleavable symmetric peptide
-
?
Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val + H2O
Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val
show the reaction diagram
-
i.e. synthetic peptide P6-P7', hydrolyzed with 55% relative efficiency compared to P8-P8'
-
?
Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
RKGDIKS-(3-nitrotyrosine)-GPGP-(lysine-anthranilide)-Y + H2O
RKGDIKS-(3-nitrotyrosine) + GPGP-(lysine-anthranilide)-Y
show the reaction diagram
-
-
-
-
?
RKGDIKSY-p-nitroanilide + H2O
RKGDIKSY + p-nitroaniline
show the reaction diagram
-
-
-
-
?
RKGDIKSYG + H2O
RKGDIKSY + glycine
show the reaction diagram
-
-
-
-
?
RKGDIKSYGLGPR + H2O
RKGDIKSY + GLGPR
show the reaction diagram
-
-
-
-
?
RKGDIKSYGLGPRYGG + H2O
RKGDIKSY + GLGPRYGG
show the reaction diagram
-
-
-
-
?
RKGDIKT-(3-nitrotyrosine)-GPGP-(lysine-anthranilide)-Y + H2O
RKGDIKT-(3-nitrotyrosine) + GPGP-(lysine-anthranilide)-Y
show the reaction diagram
-
-
-
-
?
Ser-Arg-Ala-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Ser-Arg-Ala-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
-
peptide derived from poliovirus type 1 polyprotein
-
?
Ser-Thr-Lys-Asn-Leu-Thr-Thr-Gly-Phe-Gly-His-Gln-Asn-Lys-Ala + H2O
Ser-Thr-Lys-Asn-Leu-Thr-Thr-Tyr + Gly-Phe-Gly-His-Gln-Asn-Lys-Ala
show the reaction diagram
-
synthetic hexadecapeptide corresponding to P1/P2 junction
-
?
Ser-Thr-Lys-Asp-Ile-Thr-Thr-Tyr-Gly-Phe-Gly-His-Gln-Asn-Lys-Ala + H2O
Ser-Thr-Lys-Asp-Ile-Thr-Thr-Tyr + Glys-Phe-Gly-His-Gln-Asn-Lys-Ala
show the reaction diagram
-
poor substrate
-
?
serum response factor + H2O
?
show the reaction diagram
-
-
-
-
?
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + glycine
show the reaction diagram
-
i.e. synthetic peptide P8-P1', poor substrate, smallest cleavable asymmetric peptide
-
?
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro
show the reaction diagram
-
i.e. synthetic peptide P8-P2', hydrolyzed with about the same relative efficiency as P8-P8'
-
-
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp
show the reaction diagram
-
i.e. peptide P8-P4'
-
?
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met
show the reaction diagram
-
i.e. peptide P8-P5'
-
?
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr
show the reaction diagram
-
i.e. synthetic peptide P8-6'
-
?
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val
show the reaction diagram
-
i.e. synthetic peptide P8-7'
-
?
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His + H2O
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala + Gly-Pro-Ser-Asp-Met-Tyr-Val-His
show the reaction diagram
thrombin + H2O
?
show the reaction diagram
-
-
-
?
TRPIITT-(3-nitrotyrosine)-GPSD-(lysine-anthranilate)-Y + H2O
TRPIITT-(3-nitrotyrosine) + GPSD-(lysine-anthranilate)-Y
show the reaction diagram
-
-
-
-
?
TRPIITTA-p-nitroanilide + H2O
TRPIITTA + p-nitroaniline
show the reaction diagram
-
-
-
-
?
TRPIITTAGPSDMYV + H2O
TRPIITTA + GPSDMYV
show the reaction diagram
-
-
-
-
?
additional information
?
-
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
cellular eukaryotic translation initiation factor eIF4GI + H2O
peptides
show the reaction diagram
-
-
-
-
?
cellular eukaryotic translation initiation factor eIF4GII + H2O
peptides
show the reaction diagram
-
-
-
-
?
CVB1 protein + H2O
?
show the reaction diagram
-
-
-
-
?
cytokeratin 8 + H2O
?
show the reaction diagram
dystrophin + H2O
?
show the reaction diagram
eIF4G + H2O
?
show the reaction diagram
-
eukaryotic initiation factor-4G
-
-
?
eIF4G + H2O
fragments of eIF4G
show the reaction diagram
-
-
-
-
?
eIF4GI + H2O
fragments of eIF4GI
show the reaction diagram
-
-
-
-
?
eukaryotic initiation factor 4G + H2O
?
show the reaction diagram
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
eukaryotic translation initiation factor 4G I + H2O
?
show the reaction diagram
Q9QF31
-
-
-
?
eukaryotic translation initiation factor 4G II + H2O
?
show the reaction diagram
Q9QF31
-
-
-
?
eukaryotic translation initiation factor 4gamma + H2O
?
show the reaction diagram
-
-
-
-
?
picornavirus polyprotein + H2O
?
show the reaction diagram
-
intramolecular reaction, processing begins before synthesis of the polyprotein is complete, cleavage separates capsid protein precursor and noncapsid protein precursor
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
poly(A) binding protein + H2O
fragments of poly(A) binding protein
show the reaction diagram
-
contains 1 cleavage site for 2A proteinase within the proline-rich linker domain
-
-
?
serum response factor + H2O
?
show the reaction diagram
-
-
-
-
?
thrombin + H2O
?
show the reaction diagram
Q9QF31
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaCl
-
activation, 0.15-1.25 M
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
3,4-dichloroisocoumarin
-
-
antipain
Aprotinin
-
0.015 mM, 23% inhibition
benzyloxycarbonyl-Ile-Glu-Thr-Asp(OMe)-fluoromethylketone
-
50% inhibition at 0.0077 mM, delay of self-cleavage
benzyloxycarbonyl-LSTL-fluoromethyl ketone
-
IC50: 1050 nM
benzyloxycarbonyl-LSTT-fluoromethyl ketone
-
IC50: 550 nM
benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethyl ketone
-
-
benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone
-
50% inhibition at 0.0056 mM, delay of self-cleavage
benzyloxycarbonyl-Val-Ala-Met-fluoromethyl ketone
-
-
Cd2+
-
inhibits esterase activity
chymostatin
Cl-
-
trace amounts eliminate activity
Co2+
-
0.05 mM
Elastinal
eukaryotic release factor 3
-
increasing concentrations of recombinant His-tagged eRF3 lead to partial inhibition of 2Apro-proteolytic cleavage of poly(A) binding protein that increases modestly
-
Gly-Arg-Thr-Thr-Leu-Ser-Thr-Arg-Gly-Pro-Pro-Arg-Gly-Gly-Pro-Gly
-
-
hinokitiol
-
the cleavage of the cellular eukaryotic translation initiation factor eIF4GI by 2A protease is abolished in the presence of hinokitiol, no significant amounts of eIF4GI cleavage products are found within 24 h of infection
iodoacetamide
Leupeptin
LY343813
-
0.025 mM, 61% inhibition
LY343814
-
0.025 mM, 59% inhibition
LY353350
-
0.025 mM, 67% inhibition
N,N,N',N'-tetrakis-(2-pyridylmethyl)-ethylenediamine
-
at 0.005 mM inhibitor, 50% self-processing and eIFGI cleavage are observed at 60 min, whereas the control shows 50% cleavage at 30 min and almost complete cleavage at 60 min
PABP-interacting protein 2
-
The inhibitory effect exerts by PABP-interacting protein 2 is more pronounced on 2Apro, as the lesser concentrations of PABP-interacting protein 2 that leads to partial inhibition of poly(A) binding protein cleavage by 3Cpro results in complete inhibition of 2A-pro-directed cleavage of poly(A) binding protein. 2Apro cleavage is strongly inhibited by in non-ribosome fractions, 40S and 80S ribosomes and polysome fractions.
-
pyrithione
-
the cleavage of the cellular eukaryotic translation initiation factor eIF4GI by 2A protease is abolished in the presence of pyrithione, no significant amounts of eIF4GI cleavage products are found within 24 h of infection
tosyl-L-leucine-chloromethyl ketone
tosyl-L-phenylalanine-chloromethyl ketone
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
dithiothreitol
-
higher reducing potential in the buffer activates partial cleavage of poly(A) binding protein by 2Apro, no effect on 2Apro-mediated cleavage of eIF4G
tamoxifen
-
treatment of transgenic mice with tamoxifen induces enzyme expression
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.25
Glu-Arg-Ala-Ser-Leu-Ile-Thr-Thr-Gly-Pro-Tyr-Gly-His-Gln-Ser-Gly
-
coxsackievirus B4
0.52 - 0.54
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His
0.5
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Val-Tyr
-
human rhinovirus
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00158
TRPIITTA-p-nitroanilide
Human rhinovirus sp.
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00105
benzyloxycarbonyl-LSTL-fluoromethyl ketone
coxsackievirus
-
IC50: 1050 nM
0.00055
benzyloxycarbonyl-LSTT-fluoromethyl ketone
coxsackievirus
-
IC50: 550 nM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.8
-
human rhinovirus type 2
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.6
-
esterase activity
7 - 8.5
-
broad, synthetic peptide P8-P8' as substrate
7.4
-
assay at
8
-
reaction with 2-aminobenzoic acid-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Phe(NO2)-Ala-OH
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9.2
-
about half-maximal activity at pH 6 and 9.2, synthetic peptide P8-P8' as substrate
7 - 9
-
pH 7.0: about 50% of maximal activity, pH 9.0: about 35% of maximal activity, reaction with 2-aminobenzoic acid-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Phe(NO2)-Ala-OH
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
16700
-
coxsackievirus B4, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
x-ray crystallography
monomer
-
1 * 16700, coxsackievirus B4, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
-
the enzyme is initially synthesized in an inactive form, self-processing is prerequisite for eIF4GI cleavage
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
solution structure of enzyme
-
mutant enzyme C110A, hanging drop vapor diffusion method, using 100 mM Tris-HCl (pH 8.1), 200 mM MgCl2 and 1% (w/v) polyethylene glycol 4000, at 22C
crystals grown at 4C by hanging drop vapour diffusion method
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
-
20 min, room temperature, stable in this range, 50% loss of activity at pH 3 and 10.2
648036
5
-
below, irreversible inactivation, esterase activity
648033
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2
-
t1/2 of esterase activity: more than 100 h
20
-
solubilization enhances stability at 20C
25
-
t1/2 of esterase activity: 2 h
37
-
t1/2 of esterase activity: 5 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
insoluble enzyme can be solubilized by 0.1% sarcosyl, solubilization enhances stability at 20C, soluble enzyme is unstable in dilute solution, bovine serum albumin stabilizes
-
stable to dialysis for more than 100 h at 4C
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0C, extremely unstable in crude extracts with a half-life of 2 h, 20% ethanol stabilizes, 4C, 5 mg enzyme/ml, several months
4C, in 5% v/v glycerol, long-term storage
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography
-
ammonium sulfate precipitation, IMAC resin column chromatography, and Sephacryl S-200 gel filtration
as a fusion protein
-
Coxsackievirus B3 2Apro is purified from pET-Cx2A using ion-exchange chromatography and gel filtration
-
expression in Escherichia coli
from infected HeLa cells
-
from infected HeLa cells (type S3 cells); type 1
-
glutathione-Sepharose 4B column chromatography, Ni-NTA spin column chromatography, MonoQ column chromatography, and HiLoad Superdex gel filtration
-
HeLa Ohio cells, 4-5 h after infection, to near homogeneity; type 2
-
HiTrap Q column chromatography
HRV 2; recombinant enzyme as fusion protein
-
ion-exchange chromatography (Q-Sepharose)
-
Ni-NTA resin column chromatography, Hitrap Q column chromatography, and Superdex 75 gel filtration
recombinant enzyme as expressed in Escherichia coli BL21(DE3)pLysE bearing pET8c/HRV2 2A
-
recombinant enzyme, as expressed in Escherichia coli BL21(DE3)pLysE bearing pET8c/CVB4 2A
-
recombinant protein with His-tag, in addition to main protein, detection of degradation product of 14 kDa due to cleavage at G856-V857
hepatitis A virus
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a chimeric protein (maltose-binding protein-2Apro) is expressed
-
coxsackievirus B4
-
expressed in Escherichia coli
expressed in Escherichia coli BL21 cells
-
expressed in Escherichia coli BL21(DE3)pLysS
-
expressed in Escherichia coli Rosetta cells
expressed in Escherichia coli strains Rosetta2(DE3)-pLysS and BL21-DE3 CodonPlus RILP
expressed in HeLa cells
expressed in Huh7-T7 cells
poliovirus 2A
-
expressed in Mus musculus cardiac myocytes
-
expression in Escherichia coli
-
human rhinovirus type 2
-
to test whether picornaviral 2A sequences can be used to express foreign genes in adenoviruses, 2A skipping site are inserted after the protein IX gene in an oncolytic virus that targets colon cancer cells
-
various reporter plasmids are transfected into recombinant vaccinia virus-infected BHK cells alone or with the plasmid pGEM3Z/J1, which expresses the swine vesicular disease virus 2A protease, the plasmids are transfected using FuGene6 into cells previously infected with the vaccinia virus vTF7-3, which expresses the T7 RNA polymerase
-
wild-type and mutant picornaviruses are isolated from HeLa cells transfected with in vitro-synthesized RNA using DEAE-dextran, RNA transcripts are produced from a picornavirus type 1 Mahoney P1M infectious clone, pT7M, or a picornavirus type 2 Lansing (P2L) infectious clone, pT7L
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D136N
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
D39E
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
G122E
the mutant exhibits very low cleavage efficiency toward eukaryotic initiation factor 4G
L40F
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
S67F
the mutant shows reduced cleavage efficiency toward eukaryotic initiation factor 4G
V120M
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
Y89L
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
Y90L
the mutant shows reduced cleavage efficiency toward eukaryotic initiation factor 4G
D136N
Coxsackievirus B3 Woodruff
-
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
-
D39E
Coxsackievirus B3 Woodruff
-
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
-
G122E
Coxsackievirus B3 Woodruff
-
the mutant exhibits very low cleavage efficiency toward eukaryotic initiation factor 4G
-
L40F
Coxsackievirus B3 Woodruff
-
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
-
S67F
Coxsackievirus B3 Woodruff
-
the mutant shows reduced cleavage efficiency toward eukaryotic initiation factor 4G
-
C110A
inactive
D136N
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
D144A
the mutant shows increased activity compared to the wild type enzyme
D144A/E145A
inactive
D39E
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
DELTA140-143
inactive
E145A
the mutant shows strongly reduced activity compared to the wild type enzyme
G122E
the mutant exhibits very low cleavage efficiency toward eukaryotic initiation factor 4G
L40F
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
R55A
the mutant shows reduced activity compared to the wild type enzyme
S67F
the mutant shows reduced cleavage efficiency toward eukaryotic initiation factor 4G
V120M
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
Y89L
the mutant shows wild type cleavage efficiency toward eukaryotic initiation factor 4G
Y90L
the mutant shows reduced cleavage efficiency toward eukaryotic initiation factor 4G
G60R
-
mutant is devoid of eIF4G cleavage activity
134Rstop
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
C101S
-
intramolecular cleavage is reduced to 52% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 20% of that of the wild-type enzyme, cleavage of eIF4G is 75-100% of that of the wild-type enzyme
C106S
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
C112S
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished, no detectable cleavage of eIF5G
C138A
-
intramolecular cleavage is reduced to 90% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 200% of that of the wild-type enzyme, cleavage of eIF4G is 75-100% of that of the wild-type enzyme
C52A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished, no detectable cleavage of eIF5G
C54A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished, no detectable cleavage of eIF5G
C61A
-
intramolecular cleavage is reduced to 54% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 40% of that of the wild-type enzyme, cleavage of eIF4G is 10-25% of that of the wild-type enzyme
D105N
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
D105T
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
D132R/R134D
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
D132T
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
D35A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
D35E
-
intramolecular cleavage is reduced to 16% of the wild-type activity, cleavage of P8-P8' is completely abolished
D35T
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
DELTAG104
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
DELTAG107/108
-
cleavage of P8-P8' is completely abolished
F130L
-
intramolecular cleavage is completely abolished
F130S
-
intramolecular cleavage is completely abolished
F130V
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
F136V
-
intramolecular cleavage is reduced to 57% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 25% of that of the wild-type enzyme, cleavage of eIF4G is 10-25% of that of the wild-type enzyme
G115A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
G115S
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
G118A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
G118S
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
G123A
-
intramolecular cleavage is reduced to 23% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 8% of that of the wild-type enzyme, cleavage of eIF4G is less than 10% of that of the wild-type enzyme
G123A/G124A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
G123S
-
intramolecular cleavage is reduced to 20% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 10% of that of the wild-type enzyme, cleavage of eIF4G is below 10% of that of the wild-type enzyme
G124A
-
intramolecular cleavage is reduced to 22% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 9% of that of the wild-type enzyme, cleavage of eIF4G is 10-25% of that of the wild-type enzyme
G124S
-
intramolecular cleavage is reduced to 30% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 10% of that of the wild-type enzyme, cleavage of eIF4G is 10-25% of that of the wild-type enzyme
H114G
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished, no detectable cleavage of eIF5G
H114N
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished; mutant enzyme does not contain Zn2+
H135stop
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
H18A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
H18Y
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
K113P/H114A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
L19S
-
cleavage of P8-P8' is reduced to 42% of the wild-type activity; intramolecular cleavage activity is reduced to 80% of that of the wild-type activity, cleavage of P8-P8' is reduced to 42% of the wild-type activity
N16A
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
R134Q
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished, no cleavage of eIF4G is detected
T121Y
-
intramolecular cleavage and cleavage of P8-P8' is completely abolished
Y289R
-
inactive, substitution of 2Apro A104 can partially restore self-processing on the mutant Y289R
Y289R/A104C
-
substitution of 2Apro A104 can partially restore self-processing on the mutant Y289R
Y289R/A104S
-
substitution of 2Apro A104 can partially restore self-processing on the mutant Y289R
Y85K
-
intramolecular cleavage is reduced to 82% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 39% of that of the wild-type enzyme, cleavage of eIF4G is 25-50% of that of the wild-type enzyme
Y85T
-
intramolecular cleavage is reduced to 27% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 8% of that of the wild-type enzyme, cleavage of eIF4G is 10-25% of that of the wild-type enzyme; intramolecular cleavage is reduced to 74% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 32% of that of the wild-type enzyme, cleavage of eIF4G is 25-50% of that of the wild-type enzyme
Y86F
-
intramolecular cleavage is reduced to 62% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 21% of that of the wild-type enzyme, cleavage of eIF4G is 25-50% of that of the wild-type enzyme
Y86K
-
intramolecular cleavage is reduced to 71% of that of the wild-type enzyme and cleavage of P8-P8' is reduced to 34% of that of the wild-type enzyme, cleavage of eIF4G is 25-50% of that of the wild-type enzyme
V54A
-
a BseRI fragment 4742-6148 is transferred from Se1-3C-02 DNA to pT7M, leads to defects in trans but not cis cleavage
Y88L
-
the BstEII fragment of picornavirus DNA from nucleotide 3240-3930 is replaced with a PCR product containing the appropriate mutation, the growth of picornavirus mutant is similar to that of wild-type virus in untreated cells but is completely inhibited in IFN-alpha-treated cells
Y88S
-
the BstEII fragment of picornavirus DNA from nucleotide 3240-3930 is replaced with a PCR product containing the appropriate mutation, the growth of picornavirus mutant is similar to that of wild-type virus in untreated cells but is completely inhibited in IFN-alpha-treated cells
R20D
-
significant defect in self-cleavage activity
R20G
-
significant defect in self-cleavage activity
R20P
-
complete inactivation, no self-cleavage activity
R20V
-
significant defect in self-cleavage activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
molecular biology
-
picornaviral 2A sequences can be used to express transgenes in oncolytic adenoviruses