Information on EC 3.4.22.29 - picornain 2A

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The expected taxonomic range for this enzyme is: Picornaviridae

EC NUMBER
COMMENTARY
3.4.22.29
-
RECOMMENDED NAME
GeneOntology No.
picornain 2A
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
selective cleavage of Tyr-/-Gly bond in picornavirus polyprotein
show the reaction diagram
-
-
-
-
selective cleavage of Tyr-/-Gly bond in picornavirus polyprotein
show the reaction diagram
substrate region exchanges in and out of a conformation in which it occupies the active site with association and dissociation rates in the range of 100 to 1000 per s
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
-
-
hydrolysis of peptide bond
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2A protease
-
-
-
-
2A protease
-
-
2A protease
-
-
2A protease
swine vesicular disease virus J1
-
-
-
2A proteinase
-
-
-
-
2A proteinase
-
-
2A proteinase
-
-
P2A
-
-
-
-
picornaviral 2A protein
-
-
picornaviral 2A proteinase
-
-
-
-
picornavirus 2A proteinase
-
-
-
-
picornavirus endopeptidase 2A
-
-
-
-
poliovirus 2A protease
-
-
poliovirus 2Apro
-
-
poliovirus protease 2A
-
-
-
-
poliovirus protease 2Apro
-
-
protease 2A
-
-
-
-
proteinase 2A
-
-
proteinase 2Apro
-
-
-
-
proteinase, poliovirus, 2A
-
-
-
-
rhinovirus protease 2A
-
-
-
-
Y-G proteinase 2A
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
103406-62-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
B4; types B1, A9, A21
-
-
Manually annotated by BRENDA team
coxsackievirus B3, protease 2A
-
-
Manually annotated by BRENDA team
coxsackievirus B3
B3
-
-
Manually annotated by BRENDA team
coxsackievirus B4
B4
-
-
Manually annotated by BRENDA team
bovine enterovirus
-
-
Manually annotated by BRENDA team
bovine enterovirus; bovine enterovirus type 70
-
-
Manually annotated by BRENDA team
type 1 (Mahoney strain)
-
-
Manually annotated by BRENDA team
type 1 (Mahoney strain); type 3
-
-
Manually annotated by BRENDA team
type 2 (P712-ch-2ab)
-
-
Manually annotated by BRENDA team
serotype 14
-
-
Manually annotated by BRENDA team
serotypes 14
-
-
Manually annotated by BRENDA team
serotypes 14; serotypes 2
-
-
Manually annotated by BRENDA team
serotypes 2; serotypes 2 (HRV 2)
-
-
Manually annotated by BRENDA team
swine vesicular disease virus J1
strain J1
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
a PV 2Apro variant deficient in eukaryotic initiation factor (eIF) 4GI cleavage does not increase picornavirus IRES-driven translation
physiological function
-
Poliovirus 2A protease is able to confer high translatability on picornavirus IRESs when these are transcribed from sindbis virus replicons
physiological function
-
cleavage of eIF4G by PV 2Apro in mammalian cells modifies the requirement for eIF2 in translation directed by picornavirus IRESs. Cleavage of eIF4G by PV 2Apro establishes a mechanism for IRES-driven translation that is cap- and eIF2 independent
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
-
-
-
-
?
cytokeratin 8 + H2O
?
show the reaction diagram
-
-
-
-
?
cytokeratin 8 + H2O
?
show the reaction diagram
-
the cleavage results in removal of 14 amino acids from the N-terminal head domain of cytokeratin 8, the cleavage results in removal of 14 amino acids from the N-terminal head domain of cytokeratin 8. Cleavage occurs late in the infection cycle at the time of the onset of the cytopathic effect
-
-
?
DIKSYGLGPRYGG + H2O
DIKSY + GLGPRYGG
show the reaction diagram
-
-
-
-
?
dystrophin + H2O
?
show the reaction diagram
-
human dystrophin is cleaved between residue 588 and 589 and between residue 2434 and 2435. Rat dystrophin is cleaved between resiue 590 and 591 and between residue 2427 and 2428
-
-
?
dystrophin + H2O
?
show the reaction diagram
-
cleavage of mouse dystrophin at residue 2427 and human dystrophin at residue 2434
-
-
?
dystrophin + H2O
?
show the reaction diagram
-
the protease cleaves dystrophin in coxsackievirus B3-infected myocytes. The cleavage functionally impairs dystrophin
-
-
?
dystrophin + H2O
?
show the reaction diagram
-
functional impairment and morphological disruption of dystrophin
-
-
?
dystrophin + H2O
?
show the reaction diagram
coxsackievirus B3
-
cleavage of mouse dystrophin at residue 2427 and human dystrophin at residue 2434, functional impairment and morphological disruption of dystrophin
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
-
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
-
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
efficient cleavage
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
self-processing is prerequisite for eIF4GI cleavage
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
cleavage reactions utilizing recombinant eIF4G containing a G486E substitution at the cleavage site results in drastically reduced clevage activity
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
cleavage reactions utilizing recombinant eIF4G containing a G486E substitution at the cleavage site results in drastically reduced clevage activity
-
-
-
eIF4G + H2O
?
show the reaction diagram
-
cleavage reactions utilizing recombinant eIF4G containing a G486E substitution at the cleavage site results in drastically reduced clevage activity
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
recombinant enzyme cleaves either rabbit or human eIF4G or their derived peptides, cleaves the 4G-derived peptides with 100fold lower efficiency than with a peptide derived from the poliovirus polyprotein. Up to 25fold molar excess of the enzyme over eIF4G protein is required to cause greater than 50% cleavage
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
cleavage site: PLLNV699-/-GSR
-
-
?
eIF4G + H2O
?
show the reaction diagram
-
eukaryotic initiation factor-4G
-
-
?
eIF4G + H2O
?
show the reaction diagram
coxsackievirus B4
-
cleavage reactions utilizing recombinant eIF4G containing a G486E substitution at the cleavage site results in drastically reduced clevage activity
-
-
-
eIF4G + H2O
fragments of eIF4G
show the reaction diagram
-
-
-
-
?
eIF4GI + H2O
fragments of eIF4GI
show the reaction diagram
-
-
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
-
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
in vitro
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
in vitro
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
mapping of cleavage site
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
i.e. (eIF)-4Fgamma
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
i.e. (eIF)-4Fgamma
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
from HeLa cell extracts
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
from rabbit
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
from rabbit
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
cleavage site: Arg486-Gly (rabbit), Arg485-Gly (human)
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
in-trans activity
-
-
-
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
in vivo
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
-
the 2Apro cleavage site on eIF4GI is TLSTR*GPPR
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
coxsackievirus B4
-
in vitro, i.e. (eIF)-4Fgamma, from HeLa cell extracts, from rabbit
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
protein CPa + protein CPb
show the reaction diagram
coxsackievirus B4
-
in vitro, mapping of cleavage site, i.e. (eIF)-4Fgamma, from rabbit, cleavage site: Arg486-Gly (rabbit), Arg485-Gly (human)
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
-
turns off host-cell protein synthesis
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
-
involved in shut-off in translation of cellular mRNAs upon viral infection, induces cleavage of eukaryotic initiation factor (eIF)4gamma component (i.e. p220) of eIF-4, formerly eIF-4F
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
-
involved in shut-off in translation of cellular mRNAs upon viral infection, induces cleavage of eukaryotic initiation factor (eIF)4gamma component (i.e. p220) of eIF-4, formerly eIF-4F
-
-
?
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
-
-
-
-
?
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
-
from poliovirus
-
-
-
P1/P2 precursor polypeptide + H2O
poliovirus 1C'D polypeptide + proteinase 2Apro
show the reaction diagram
-
from poliovirus
-
-
?
P1/P2 precursor polypeptide + H2O
poliovirus 1C'D polypeptide + proteinase 2Apro
show the reaction diagram
-
from poliovirus
-
-
-
P1/P2 precursor polypeptide + H2O
poliovirus 1C'D polypeptide + proteinase 2Apro
show the reaction diagram
-
from poliovirus
-
-
?
P1/P2 precursor polypeptide + H2O
poliovirus 1C'D polypeptide + proteinase 2Apro
show the reaction diagram
-
from rhinovirus, cleaves P1/P2 junction
-
-
?
P8-P8' + H2O
?
show the reaction diagram
-
-
-
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
-
-
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
-
-
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
immediately after protease 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent rhinovirus or enterovirus chain is cleaved intramolecularly
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleavage between C-terminus of VP1 and N-terminus of picornain 2A
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves intermolecularly at a second site in the 3D protein between 3C' and 3D'
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
-
cleaves Tyr-Gly pairs in both Thr-Tyr-Gly sequences of poliovirus polyprotein, cleaves Thr-Gly in Thr-Thr-Gly (serotype B1) or Asn-Thr-Gly (serotype B3) of coxsackievirus polyprotein, cleaves Ala-Gly in Thr-Ala-Gly (HRV 2), Tyr-Gly in Ser-Tyr-Gly (HRV 14) and Val-Gly in Asn-Val-Gly (HRV 89) in rhinovirus polyproteins
-
?
picornavirus polyprotein + H2O
hydrolyzed picornavirus polyprotein
show the reaction diagram
coxsackievirus B4
-
-
-
-
?
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
-
involved in primary processing of poliovirus
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
-
involved in primary processing of poliovirus
-
-
-
poliovirus polypeptide + H2O
?
show the reaction diagram
-
immediately after polypeptide 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent chain is cleaved intramolecularly
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
-
together with 3C protease
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
coxsackievirus B4
-
involved in primary processing of poliovirus
-
-
-
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
-
-
-
-
?
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
-
i.e. synthetic peptide P6-P8', hydrolyzed with 70% relative efficiency compared to P8-P8'
-
?
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
-
?
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
-
-
-
-
-
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
-
-
-
?
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
-
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
i.e. synthetic peptide P8-P8'
-
?
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
-
best substrate for rhinovirus enzyme
-
-
?
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
coxsackievirus B4
-
-
-
?
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
-
-
-
-
?
modified pentadecameric peptides + H2O
?
show the reaction diagram
-
i.e. synthetic P8-P7' peptides, intermolecular specificity, changes at P2 and P1' are highly deleterious
-
-
?
additional information
?
-
-
-
-
-
-
additional information
?
-
-
cleavage specificity
-
-
-
additional information
?
-
-
cleavage specificity
-
-
-
additional information
?
-
-
cleavage specificity
-
-
-
additional information
?
-
-
cleavage specificity
-
-
-
additional information
?
-
-
cleavage specificity
-
-
-
additional information
?
-
-
cleavage specificity
-
-
-
additional information
?
-
-
the enzyme displays a charged aspartic acid, which may significantly influence the interaction between the thiol and imidazole groups
-
-
-
additional information
?
-
-
no hydrolysis of some modified pentadecameric synthetic peptides P8-P7'(overview), synthetic peptides P3-P8', Arg-Lys-Gly-Asp-Ile-Lys-Ser-Tyr-Gly-Ile-Gly-Pro-Arg-Tyr-Gly-Gly, Asn-Val-Arg-Ala-Val-Lys-Asn-Val-Gly-Pro-Ser-Asp-Met-Tyr-Val-His, Asp-Val-Phe-Thr-Asn-Val-Gly-Pro-Ser-Ser-Met-Phe-Val-His
-
-
-
additional information
?
-
-
no hydrolysis of some modified pentadecameric synthetic peptides P8-P7'(overview), synthetic peptides P3-P8', Arg-Lys-Gly-Asp-Ile-Lys-Ser-Tyr-Gly-Ile-Gly-Pro-Arg-Tyr-Gly-Gly, Asn-Val-Arg-Ala-Val-Lys-Asn-Val-Gly-Pro-Ser-Asp-Met-Tyr-Val-His, Asp-Val-Phe-Thr-Asn-Val-Gly-Pro-Ser-Ser-Met-Phe-Val-His
-
-
-
additional information
?
-
-
no cleavage by single-site mutant enzymes with substitutions at Cys55, Cys57, Cys115 or His117
-
-
-
additional information
?
-
-
synthetic peptides with replacements of P1 or P1' residues with epsilon-amino caproic acid or statine
-
-
-
additional information
?
-
-
residues Gly123, Gly124, Thr121 and Cys101 are proposed to be involved in the architecture of the substrate binding pocket and to provide the correct environment for the catalytic triad of His18, Asp35, and Cys106. Residues Tyr85 and Tyr86 are thought to participate in substrate recognition
-
-
-
additional information
?
-
-
the primary cleavage in cardio- and aphthovirus occurs between 2A and 2B (cleavage site: Gly-Pro in-Asn-Pro-Gly-Pro-)
-
-
-
additional information
?
-
-
the primary cleavage in cardio- and aphthovirus occurs between 2A and 2B (cleavage site: Gly-Pro in-Asn-Pro-Gly-Pro-)
-
-
-
additional information
?
-
-
the primary cleavage in cardio- and aphthovirus occurs between 2A and 2B (cleavage site: Gly-Pro in-Asn-Pro-Gly-Pro-)
-
-
-
additional information
?
-
-
the primary cleavage in cardio- and aphthovirus occurs between 2A and 2B (cleavage site: Gly-Pro in-Asn-Pro-Gly-Pro-)
-
-
-
additional information
?
-
-
the primary cleavage in cardio- and aphthovirus occurs between 2A and 2B (cleavage site: Gly-Pro in-Asn-Pro-Gly-Pro-)
-
-
-
additional information
?
-
-
enzyme is active both as structural element of a precursor and as mature protein
-
-
-
additional information
?
-
-
the active site of the enzyme consists of a catalytic triad formed by His18, Asp35 and Cys106
-
-
-
additional information
?
-
-
enzyme, or its polyprotein precursors, are required to stabilize viral RNA and prolong translation. Enzyme activity stimulates negative-strand initiation by approximately fivefold, but has no effect on stability. Stimulation of negative-strand synthesis is independent of the effect on stability and translation
-
-
-
additional information
?
-
-
does not cleave death-associated protein 5
-
-
-
additional information
?
-
-
fusion of the C-terminal three amino acid residues of VP1 to the N-terminus of 2Apro is necessary for specific self-cleavage activity to generate mature 2Apro
-
-
-
additional information
?
-
-
2Apro cleaves eIF4G, separating the domains of eIF4GI or eIF4GII that bind to eIF4E (and mRNA) and eIF3 (and the 40S ribosome) and inhibits de novo translation initiation by interfering with the ribosome mRNA binding step
-
-
-
additional information
?
-
-
2Apro processes the viral polyprotein, and it cleaves a variety of host proteins, including the translation proteins eIF4GI, eIF4GII and poly(A)-binding protein, important cellular substrate for 2Apro is eIF4G, cleavage of this translation protein leads to inhibition of host protein synthesis during picornavirus infection, inhibitory role for 2Apro in the most downstream event in interferon signaling, the antiviral activities of interferon-stimulated genes
-
-
-
additional information
?
-
-
picornaviral 2A sequences are used to express transgenes in oncolytic adenoviruses
-
-
-
additional information
?
-
-
proteinase 2Apro is essential for enterovirus replication in type I interferon-treated cells
-
-
-
additional information
?
-
-
ribosome skipping is a mechanism that allows translation of multiple proteins from a single mRNA, it is based on the use of a picornaviral 2A sequence that causes the ribosome to continue translation after skipping the formation of one peptide bond
-
-
-
additional information
?
-
coxsackievirus B4
-
cleavage specificity
-
-
-
additional information
?
-
coxsackievirus B4
-
-
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
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
-
-
-
-
?
cytokeratin 8 + H2O
?
show the reaction diagram
-
-
-
-
?
cytokeratin 8 + H2O
?
show the reaction diagram
-
the cleavage results in removal of 14 amino acids from the N-terminal head domain of cytokeratin 8. Cleavage occurs late in the infection cycle at the time of the onset of the cytopathic effect
-
-
?
dystrophin + H2O
?
show the reaction diagram
-
the protease cleaves dystrophin in coxsackievirus B3-infected myocytes. The cleavage functionally impairs dystrophin
-
-
?
dystrophin + H2O
?
show the reaction diagram
coxsackievirus, coxsackievirus B3
-
functional impairment and morphological disruption of dystrophin
-
-
?
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 translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
-
turns off host-cell protein synthesis
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
-
involved in shut-off in translation of cellular mRNAs upon viral infection, induces cleavage of eukaryotic initiation factor (eIF)4gamma component (i.e. p220) of eIF-4, formerly eIF-4F
-
-
?
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
-
involved in primary processing of poliovirus
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
-
involved in primary processing of poliovirus
-
-
-
poliovirus polypeptide + H2O
?
show the reaction diagram
-
immediately after polypeptide 2A has been synthesized the Tyr-Gly pair between P1 and P2 of the nascent chain is cleaved intramolecularly
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
-
together with 3C protease
-
-
?
poliovirus polypeptide + H2O
?
show the reaction diagram
coxsackievirus B4
-
involved in primary processing of poliovirus
-
-
-
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
-
-
?
eukaryotic translation initiation factor 4F p220 subunit + H2O
?
show the reaction diagram
-
involved in shut-off in translation of cellular mRNAs upon viral infection, induces cleavage of eukaryotic initiation factor (eIF)4gamma component (i.e. p220) of eIF-4, formerly eIF-4F
-
-
?
additional information
?
-
-
enzyme, or its polyprotein precursors, are required to stabilize viral RNA and prolong translation. Enzyme activity stimulates negative-strand initiation by approximately fivefold, but has no effect on stability. Stimulation of negative-strand synthesis is independent of the effect on stability and translation
-
-
-
additional information
?
-
-
2Apro cleaves eIF4G, separating the domains of eIF4GI or eIF4GII that bind to eIF4E (and mRNA) and eIF3 (and the 40S ribosome) and inhibits de novo translation initiation by interfering with the ribosome mRNA binding step
-
-
-
additional information
?
-
-
2Apro processes the viral polyprotein, and it cleaves a variety of host proteins, including the translation proteins eIF4GI, eIF4GII and poly(A)-binding protein, important cellular substrate for 2Apro is eIF4G, cleavage of this translation protein leads to inhibition of host protein synthesis during picornavirus infection, inhibitory role for 2Apro in the most downstream event in interferon signaling, the antiviral activities of interferon-stimulated genes
-
-
-
additional information
?
-
-
picornaviral 2A sequences are used to express transgenes in oncolytic adenoviruses
-
-
-
additional information
?
-
-
proteinase 2Apro is essential for enterovirus replication in type I interferon-treated cells
-
-
-
additional information
?
-
-
ribosome skipping is a mechanism that allows translation of multiple proteins from a single mRNA, it is based on the use of a picornaviral 2A sequence that causes the ribosome to continue translation after skipping the formation of one peptide bond
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
NaCl
-
activation, 0.15-1.25 M
Zn2+
-
zinc-containing cysteine proteinase, atom emission spectroscopy: 1 mol/mol enzyme; Zn2+-depleted enzyme is reactivated by Zn2+, not Co2+ or Ni2+; Zn2+ is not involved in catalysis but is required for generation of an active enzyme
Zn2+
-
no mechanistic involvement; Zn2+-binding motif
Zn2+
-
the enzyme contains a structurally important zinc ion
Zn2+
-
a tightly bound Zn2+ is essential for stability of the enzyme, is tetrahedrally coordinated by three cysteine sulfurs and one histidine nitrogen
Zn2+
-
the enzyme contains a tightly bound Zn2+ required for structural integrity; the zinc-binding motif comprises Cys52, Cys54, Cys112 and His114
additional information
-
human rhinovirus HRV 2 enzyme is largely unaffected by ionic strength
additional information
-
-
additional information
-
no activation by eukaryotic elongation initiation factor 3, eIF-3
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1,10-phenanthroline
-
in trans cleavage activity
1,10-phenanthroline
-
-
3,4-Dichloroisocoumarin
-
-
antipain
-
eIF-4gamma as substrate in HeLa cell extracts; human rhinovirus
antipain
-
peptide or eIF cleavage
antipain
-
human rhinovirus; peptide or eIF cleavage
Aprotinin
-
0.015 mM, 23% inhibition
benzyloxycarbonyl-Ile-Glu-Thr-Asp(OMe)-fluoromethylketone
-
50% inhibiton 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% inhibiton at 0.0056 mM, delay of self-cleavage
benzyloxycarbonyl-Val-Ala-Met-fluoromethyl ketone
-
-
Ca2+
-
not (10 mM, esterase activity)
Ca2+
-
50 mM, weak
Cd2+
-
inhibits esterase activity
chymostatin
-
eIF-4gamma as substrate in HeLa cell extracts; human rhinovirus
chymostatin
-
human rhinovirus
Cl-
-
trace amounts eliminate activity
Cu2+
-
weak, 1 mM, inhibits esterase activity
EDTA
-
at high concentration
Elastinal
-
eIF-4gamma as substrate in HeLa cell extracts; human rhinovirus
Elastinal
-
human rhinovirus
Elastinal
-
0.1 mM, 68% inhibition
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
-
inhibits esterase activity
iodoacetamide
-
strong
iodoacetamide
-
1 mM, complete inhibition
Leupeptin
-
inhibits esterase activity
LY343813
-
0.025 mM, 61% inhibition
LY343814
-
0.025 mM, 59% inhibition
LY353350
-
0.025 mM, 67% inhibition
Mg2+
-
not (10 mM, esterase activity)
Mg2+
-
50 mM, weak
Mn2+
-
10 mM, inhibits esterase activity; weak
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
NEM
-
inhibits esterase activity
NEM
-
1 mM, complete inhibition
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
-
inhibits esterase activity
tosyl-L-leucine-chloromethyl ketone
-
-
tosyl-L-leucine-chloromethyl ketone
-
1 mM, complete inhibition
tosyl-L-phenylalanine-chloromethyl ketone
-
inhibits esterase activity
tosyl-L-phenylalanine-chloromethyl ketone
-
weak
Zn2+
-
1 mM, inhibits esterase activity
additional information
-
no inhibition by trans-epoxysuccinyl-L-leucylamido(4-guanidino)-butane (i.e. E-64)
-
additional information
-
no inhibition by pepstatin (esterase activity); no inhibition by PMSF (esterase activity)
-
additional information
-
-
-
additional information
-
no inhibition by 1,7-phenanthroline and DTT; no inhibition by EGTA
-
additional information
-
-
-
additional information
-
no inhibition by amastatin, bestatin, epiamastatin, foroxymithin, phosphoramidon, Nle-statine-Ala-statine and APMSF; no inhibition by trans-epoxysuccinyl-L-leucylamido(4-guanidino)-butane (i.e. E-64)
-
additional information
-
no inhibition by trans-epoxysuccinyl-L-leucylamido(4-guanidino)-butane (i.e. E-64)
-
additional information
-
metal-chelating agent
-
additional information
-
no inhibition by pepstatin
-
additional information
-
no inhibition by unmethylated benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone or benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethylketone
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.25
-
Glu-Arg-Ala-Ser-Leu-Ile-Thr-Thr-Gly-Pro-Tyr-Gly-His-Gln-Ser-Gly
-
coxsackievirus B4
0.52
-
Thr-Arg-Pro-Ile-Ile-Thr-Thr-Ala-Gly-Pro-Ser-Asp-Met-Tyr-Val-His
-
human rhinovirus type 2
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]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00158
-
TRPIITTA-p-nitroanilide
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00105
-
benzyloxycarbonyl-LSTL-fluoromethyl ketone
-
IC50: 1050 nM
0.00055
-
benzyloxycarbonyl-LSTT-fluoromethyl ketone
-
IC50: 550 nM
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8.8
-
-
human rhinovirus type 2
additional information
-
-
cleavage of poly(A)-binding protein by 2Apro is incomplete, a single 2Apro cleavage product can be observed late in the reaction, and 8fold-higher levels of 2A protease produce only marginally higher levels of cleavage; combined 2Apro and 3Cpro result in a net stimulation of poliovirus internal ribosome entry site-mediated translation, the resulting rate of translation is about 3fold greater than the control value but not as great as the 4fold stimulation of translation from preincubation with 2Apro alone; low concentrations of 2Apro produce complete cleavage of eIF4GI in less than 5 min, producing several cleavage products from the multiple eIF4GI isoforms
additional information
-
-
around 50% cleavage of cellular eukaryotic translation initiation factor eIF4GI is obtained at 4 h postinfection; at 6 h and 8 h postinfection, cellular eukaryotic translation initiation factor eIF4GI is nearly completely processed into its typical cleavage products
additional information
-
-
in the presence of the 2A protease, the Rluc expression from the mutant encephalomyocarditis virus internal ribosome entry site elements is strongly reduced suggesting that the residual translation (seen in the absence of 2A) is occurring principally by a cap-dependent mechanism; monocistronic RNAs containing the avian encephalomyelitis virus IRES elements with mutations in the stem 1 or stem 2 of the pseudoknot are also very efficiently translated when assayed alone, but this expression is essentially eliminated in the presence of 2A protease; when a dicistronic vector containing the Fluc coding sequence upstream of the wild type encephalomyocarditis virus internal ribosome entry site elements is assayed, it is shown that the encephalomyocarditis virus IRES-directed Rluc expression is unaffected by the coexpression of the 2A protease, but the expression of the upstream open reading frame is strongly inhibited
additional information
-
-
addition of increasing concentrations of PABP-interacting protein 1 do not affect 2Apro-mediated cleavage of poly(A) binding protein, at the highest concentration (3 microg) of PABP-interacting protein 1 tested, increasing concentrations of 2Apro (0.5-1.5 microg) lead to a dose-dependent increase in cleavage of poly(A) binding protein by 2Apro proteinase; cleavage kinetics analysis indicates that poly(A) binding protein exists in multiple conformations, some of which are resistant to 2Apro cleavage and can be modulated by reducing potential; poly(A) binding protein in a HeLa S10 lysate is also highly resistant to cleavage by 2Apro despite high 2Apro activity versus eIF4GI, the initial cleavage rate is very slow and remains constant for 60 min before slowing down further upon extended incubation. Extended incubation also reveals a biphasic cleavage profile, incubation of 2Apro with recombinant His-poly(A) binding protein results in relatively poor and variable cleavage ranging from 0-20%, this suggested that most purified protein usually exists in a conformation not suitable for 2Apro recognition and binding
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
infected Escherichia coli, containing plasmid pATH-2A
Manually annotated by BRENDA team
-
HeLa cells are infected
Manually annotated by BRENDA team
additional information
-
synthesized in cells following infection by picornaviruses
Manually annotated by BRENDA team
additional information
coxsackievirus B4
-
synthesized in cells following infection by picornaviruses
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
coxsackievirus B4
-
-
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
16700
-
-
coxsackievirus B4, gel filtration
17000
-
-
coxsackievirus B4, gel filtration
17000
-
-
-
17000
-
-
human rhinovirus type 2, gel filtration in 0.05 M Tris, 1 mM EDTA, 5 mM DTT, 5% glycerol, with 0.5 M NaCl at pH 5.9
30000
-
-
human rhinovirus HRV 2, gel filtration
30000
-
-
human rhinovirus type 2, gel filtration in 0.05 M Tris, 1 mM EDTA, 5 mM DTT, 5% glycerol, with 0.05 M NaCl at pH 8, behaving as a dimer
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 17000, SDS-PAGE
?
-
x * 16300, SDS-PAGE
?
-
x * 16000, SDS-PAGE
?
-
x * 21000, SDS-PAGE
monomer
-
1 * 16700, coxsackievirus B4, SDS-PAGE
monomer
coxsackievirus B4
-
1 * 16700, coxsackievirus B4, SDS-PAGE
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
-
the enzyme is initially synthesized in an inactive form, self-processing is prerequisite for eIF4GI cleavage
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
solution structure of enzyme
-
crystals grown at 4C by hanging drop vapour diffusion method
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
9
-
20 min, room temperature, stable in this range, 50% loss of activity at pH 3 and 10.2
5
-
-
below, irreversible inactivation, esterase activity
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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 ACCESSION NO.
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
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
0C, extremely unstable in crude extracts with a half-life of 2 h, 20% ethanol stabilizes, 4C, 5 mg enzyme/ml, several months
-
0C, extremely unstable in crude extracts with a half-life of 2 h, 20% ethanol stabilizes, 4C, 5 mg enzyme/ml, several months
-
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
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
as a fusion protein
-
Coxsackievirus B3 2Apro is purified from pET-Cx2A using ion-exchange chromatography and gel filtration
-
expression in Escherichia coli
-
glutathione-Sepharose 4B column chromatography, Ni-NTA spin column chromatography, MonoQ column chromatography, and HiLoad Superdex gel filtration
-
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
-
ion-exchange chromatography (Q-Sepharose)
-
affinity chromatography
-
expression in Escherichia coli
-
from infected HeLa cells
-
from infected HeLa cells (type S3 cells); type 1
-
HeLa Ohio cells, 4-5 h after infection, to near homogeneity; type 2
-
HRV 2; recombinant enzyme as fusion protein
-
recombinant enzyme as expressed in Escherichia coli BL21(DE3)pLysE bearing pET8c/HRV2 2A
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
coxsackievirus B4
-
expressed in Escherichia coli BL21 cells
-
expressed in HeLa cells
-
expressed in HeLa cells
-
expressed in Mus musculus cardiac myocytes
-
expressed in Escherichia coli BL21(DE3)pLysS
-
expressed in HeLa cells
-
a chimeric protein (maltose-binding protein-2Apro) is expressed
-
expressed in HeLa cells
-
expression in Escherichia coli
-
human rhinovirus type 2
-
expressed in HeLa cells
-
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
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
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
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
R20D
-
significant defect in self-cleavage activity
R20G
-
significant defect in self-cleavage activity
R20P
-
complete inactivation, no self-cleavage activity
additional information
-
recombinant protein with His-tag, no enzymic activity upon eukaryotic initiation factor 4G I
G60R
-
mutant is devoid of eIF4G cleavage activity
additional information
-
stable expression of enzyme in HeLa cell lines. Expression of enzyme leads to apoptosis, indicated by nuclear fragmentation, DNA breakdown and phosphatidylserine translocation. Enzyme induces the cleavage of poly-ADP-ribose-polymerase, which is blocked by the caspase-3 inhibitor peptide Asp-Glu-Val-Asp
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
additional information
-
the presence of an extensive C-terminal helix, in which Asp132, Arg134, Phe130 and Phe136 play important roles, explains why mutations in this region are generally detrimental to proteinase activity.The zinc-binding motif comprises Cys52, Cys54, Cys112 and His114. Exchange of these residues inactivates the enzyme
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
additional information
-
modification of R20 to every amino acid possible
additional information
-
substitution mutants of position R20 produce less severe disease
R20V
-
significant defect in self-cleavage activity
additional information
swine vesicular disease virus J1
-
substitution mutants of position R20 produce less severe disease
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
molecular biology
-
picornaviral 2A sequences can be used to express transgenes in oncolytic adenoviruses