Information on EC 3.4.22.66 - calicivirin

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The expected taxonomic range for this enzyme is: ssRNA positive-strand viruses, no DNA stage

EC NUMBER
COMMENTARY
3.4.22.66
-
RECOMMENDED NAME
GeneOntology No.
calicivirin
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
endopeptidase with a preference for cleavage when the P1 position is occupied by Glu-/- and the P1' position is occupied by Gly-/-
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3C-like protease
-
-
3C-like protease
-
-
3C-like protease
-
-
3C-like protease
norovirus MD145
-
-
-
3C-like proteinase
Q66914
-
3C-like proteinase
-
-
3C-like proteinase
norovirus MD145-12
-
-
-
3C-like viral protease
-
-
-
-
3CL Pro
-
-
3Cpro
-
-
-
-
C37.001
-
-
-
-
FCV 3CLpro
Q66914
-
NoV 3CLpro
-
-
NoV 3CLpro
norovirus MD145-12
-
-
-
NV protease
-
-
NVPro
-
-
Southampton virus 3C-like protease
-
-
-
-
viral cysteine protease
-
-
virus-encoded 3C-like proteinase
-
-
CAS REGISTRY NUMBER
COMMENTARY
218925-73-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain MD145
-
-
Manually annotated by BRENDA team
strain MD145-12
-
-
Manually annotated by BRENDA team
norovirus MD145
strain MD145
-
-
Manually annotated by BRENDA team
norovirus MD145-12
strain MD145-12
-
-
Manually annotated by BRENDA team
serotype 3
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
the enzyme is required for viral polyprotein processing
physiological function
-
the Calicivirus proteases cleaves the viral precursor polyprotein encoded by open reading frame1 into multiple intermediate and mature proteins. The Calicivirus protease is a Cys protease with catalytic Cys122, and the His39-Glu60-Cys122 catalytic triad formation is important for protease activity. The substrate recognition mechanism may be different between Caliciviridae, i.e. the Rabbit hemorrhagic disease virus and Sapporo virus proteases and the Norwalk virus and Feline calicivirus proteases. Proteolytic cleavage occurs at several cleavage sites in the ORF1 polyprotein without a functional acid residue in the FCV protease
physiological function
-
the Calicivirus proteases cleaves the viral precursor polyprotein encoded by open reading frame1 into multiple intermediate and mature proteins. The Calicivirus protease is a Cys protease with catalytic Cys139, and the His30-Glu54-Cys139 catalytic triad formation is important for protease activity. The substrate recognition mechanism may be different between Caliciviridae, i.e. the Rabbit hemorrhagic disease virus and Sapporo virus proteases and the Norwalk virus and Feline calicivirus proteases. Proteolytic cleavage occurs at several cleavage sites in the ORF1 polyprotein without a functional acid residue in the NoV protease
physiological function
-
the Calicivirus proteases cleaves the viral precursor polyprotein encoded by open reading frame1 into multiple intermediate and mature proteins. The Calicivirus protease is a Cys protease with catalytic Cys104, and the His27-Asp44-Cys104 catalytic triad formation is important for protease activity. The substrate recognition mechanism may be different between Caliciviridae, i.e. the Rabbit hemorrhagic disease virus and Sapporo virus proteases and the Norwalk virus and Feline calicivirus proteases. RHDV protease critically needs the acidic residue during catalysis
physiological function
-
the Calicivirus proteases cleaves the viral precursor polyprotein encoded by open reading frame1 into multiple intermediate and mature proteins. The Calicivirus protease is a Cys protease with catalytic Cys116, and the His31-Glu52-Cys116 catalytic triad formation is important for protease activity. The substrate recognition mechanism may be different between Caliciviridae, i.e. the Rabbit hemorrhagic disease virus and Sapporo virus proteases and the Norwalk virus and Feline calicivirus proteases. SaV protease critically needs the acidic residue during catalysis
additional information
-
acidic amino acid (Glu or Asp), as well as the His and Cys in the putative catalytic triad, cannot be replaced by Ala for normal processing activity of the ORF1 polyprotein in vitro. Similarly, normal activity is not retained if the nucleophile Cys is replaced with Ser
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
dabcyl-KTSAVLQSGFRKME-Edans + H2O
?
show the reaction diagram
-
-
-
-
?
Glu-Thr-Thr-Leu-Glu-Gly-Gly-Asp
Glu-Thr-Thr-Leu-Glu + Gly-Gly-Asp
show the reaction diagram
-
model of an oligopeptide bound to CVP
-
-
?
Murine norovirus nonstructural polyprotein + H2O
?
show the reaction diagram
-
clevage map
-
-
?
norovirus polyprotein + H2O
hydrolyzed norovirus polyprotein
show the reaction diagram
-
-
-
-
?
ORF1 polyprotein + H2O
nonstructural proteins
show the reaction diagram
-
-
-
-
?
ORF1 polyprotein + H2O
precursor comprised of both the proteinase and polymerase + mature polymerase
show the reaction diagram
norovirus, norovirus MD145
-
-
-
-
?
ORF1 polyprotein + H2O
six nonstructural proteins for replication
show the reaction diagram
Q83883
-
-
-
?
poly(A)-binding protein
?
show the reaction diagram
Human enterovirus C, norovirus, norovirus MD145-12
-
cleavage separates the C-terminal domain of the substrate that binds translation factors eIF4B and eRF3 from the N-terminal RNA-binding domain
-
-
?
Poly(A)-binding protein + H2O
?
show the reaction diagram
Q66914
cleavage separates the C-terminal domain of the substrate that binds translation factors eIF4B and eRF3 from the N-terminal RNA-binding domain
-
-
?
Southampton viral polyprotein precursor p113 + H2O
p48 N-terminal + p41 helicase + 22000 Da 3A-like protein + 16000 Da Vpg + 19000 Da 3C-like viral protease + 57000 Da 3D polymerase
show the reaction diagram
-
cleavage sites are Q399-/-G400, Q762-/-G763, E961-/-G962, E1099-/-A1100 and E1280-/-G1281
-
?
His3Cd-GST-protein + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
four amino acid residues are essential to protease activities: H31, E52, C116, and H131. These amino acids are involved in the formation of a conserved catalytic surface
-
-
-
additional information
?
-
-
four amino acid residues are essential to protease activities: H39, E60, C122, and H137. These amino acids are involved in the formation of a conserved catalytic surface
-
-
-
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
norovirus polyprotein + H2O
hydrolyzed norovirus polyprotein
show the reaction diagram
-
-
-
-
?
Southampton viral polyprotein precursor p113 + H2O
p48 N-terminal + p41 helicase + 22000 Da 3A-like protein + 16000 Da Vpg + 19000 Da 3C-like viral protease + 57000 Da 3D polymerase
show the reaction diagram
-
cleavage sites are Q399-/-G400, Q762-/-G763, E961-/-G962, E1099-/-A1100 and E1280-/-G1281
-
?
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
20% inhibited by 0.01 mM, 40% inhibited by 5 mM
4-(2-aminoethyl)-benzene-sulfonyl fluoride
Q83883
irreversible inhibition, soluble stable nontoxic alternative to phenylmethylsulfonyl fluoride
Hg2+
-
5 mM completely inhibits
K+
-
inhibited by 100 mM
L-Leu-Gly-Gly
-
-
N-ethylmaleimide
-
almost complete inhibition
N-Phenylmaleimide
-
almost complete inhibition
Na+
-
83% inhibited by 100 mM
p-chloromercuribenzenesulfonate
-
27% inhibited by 0.01 mM
p-chloromercuribenzoic acid
-
82% inhibited by 0.01 mM
phenyl methylsulfonylfluoride
-
20% inhibition by 0.01 mM
phenylmethylsulfonyl fluoride
Q83883
-
zinc N-ethyl-N-phenyldithiocarbamate
-
-
Zn2+
-
5 mM completely inhibits
methyl methanethiosulfonate
-
almost complete inhibition
additional information
-
not inhibited by Mg2+, Ca2+, 0.01 mM iodoacetate nor 0.01 mM iodoacetamide
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Glu 54
Q83883
active role in proteolysis
-
IPTG
-
induction of gene expression
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8.62
-
-
100% enzyme activity
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.22
9.08
-
-
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
80% enzyme activity
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
48
-
-
PDB
SCOP
CATH
ORGANISM
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Norwalk virus (strain GI/Human/United States/Norwalk/1968)
Southampton virus (strain GI/Human/United Kingdom/Southampton/1991)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
19000
-
-
calculation from nucleotide sequence
20600
-
-
calculated from ProHis protein
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
Q83883
size exclusion chromatography results
homodimer
-
x-ray crystallography
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapor diffusion grown at 20°C, at 2.8 A resolution
-
sitting drop vapor diffusion method, using 18% (w/v) PEG 6000, 10% (v/v) DMSO, 14% methyl-2,4-pentanediol, and 0.12 M Tris-HCl, pH 8.5
-
by hanging drop vapor diffusion or sitting drop vapor diffusion at room temperature, to 1.5 and 2.2 A resolution
Q83883
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
GSTrap FF column chromatography
-
purified over chitin beads and by gel filtration
-
Ni-NTA column chromatography and HiTrap 26/10 QFF column chroamtography
-
by nickel affinity chromatography or size exclusion chromatography
Q83883
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli BL21-CodonPlus(DE3)-RIPL cells
-
expression in Escherichia coli BL21 (DE3)
-
expression in Escherichia coli BL21 (DE3)
Q66914
expressed in Escherichia coli BL21 cells
-
expression in HeLa cells
-
cloned into pET41 Ek/LIC and pET46 Ek/LIC and expressed in Escherichia coli BL21 (DE3)
Q83883
expression in HEK293 cells
-
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C139A
-
cysteine residue responsible for hydrolysis
E54L
-
the E54L mutant protease is a substrate-specificity mutant and requires large hydrophobic amino acid residues at both P4 and P2 positions of the substrate, it cleaves the sequence 133LSFE/AP between the 3B and 3C regions of norovirus polyprotein, but does not cleaves the sequence 198ATSE/GK between the 3A and 3B
C139S
-
site-directed mutagenesis, the mutant is affected in its catalytic activity of proteolytic processing
C122S
-
site-directed mutagenesis, the mutant is affected in its catalytic activity of proteolytic processing
additional information
-
acidic amino acid (Glu or Asp), as well as the His and Cys in the putative catalytic triad, cannot be replaced by Ala for normal processing activity of the ORF1 polyprotein in vitro. Similarly, normal activity is not retained if the nucleophile Cys is replaced with Ser
E54A
Q83883
Glu at the active centre
additional information
-
acidic amino acid (Glu or Asp), as well as the His and Cys in the putative catalytic triad, cannot be replaced by Ala for normal processing activity of the ORF1 polyprotein in vitro. Similarly, normal activity is not retained if the nucleophile Cys is replaced with Ser
C104S
-
site-directed mutagenesis, the mutant is affected in its catalytic activity of proteolytic processing
additional information
-
acidic amino acid (Glu or Asp), as well as the His and Cys in the putative catalytic triad, cannot be replaced by Ala for normal processing activity of the ORF1 polyprotein in vitro. Similarly, normal activity is not retained if the nucleophile Cys is replaced with Ser
C116S
-
site-directed mutagenesis, the mutant is affected in its catalytic activity of proteolytic processing
additional information
-
acidic amino acid (Glu or Asp), as well as the His and Cys in the putative catalytic triad, cannot be replaced by Ala for normal processing activity of the ORF1 polyprotein in vitro. Similarly, normal activity is not retained if the nucleophile Cys is replaced with Ser
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
nonbacterial acute gastroenteritis and other diseases associated with viruses expressing 3Cpro, knowledge of the structure and previous mutagenesis study allows to correlate proteolysis and structure
medicine
-
poliovirus 3Cpro mediates the cleavage of poly(A)-binding protein as part of its strategy to inhibit cellular translation
medicine
-
calicivirus 3CLpro mediates the cleavage of poly(A)-binding protein as part of its strategy to inhibit cellular translation
medicine
-
norovirus precursor comprised of both the proteinase and polymerase (released from 3C-like proteinase) is a bifunctional enzyme during virus replication, which might be useful in the development of antiviral drugs of the noroviruses associated with acute gastroenteritis
medicine
norovirus MD145
-
norovirus precursor comprised of both the proteinase and polymerase (released from 3C-like proteinase) is a bifunctional enzyme during virus replication, which might be useful in the development of antiviral drugs of the noroviruses associated with acute gastroenteritis
-
medicine
norovirus MD145-12
-
calicivirus 3CLpro mediates the cleavage of poly(A)-binding protein as part of its strategy to inhibit cellular translation
-
medicine
Q83883
Norwalk virus is the major cause of acute, epidemic, viral gastroenteritis
medicine
-
establishment of a mammalian cell-based system for analysis of human norovirus replication and, thus makes it feasible to investigate antiviral agents in mammalian cells