Information on EC 3.4.22.44 - nuclear-inclusion-a endopeptidase

New: Word Map on EC 3.4.22.44
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Potyvirus

EC NUMBER
COMMENTARY hide
3.4.22.44
-
RECOMMENDED NAME
GeneOntology No.
nuclear-inclusion-a endopeptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolyses glutaminyl bonds, and activity is further restricted by preferences for the amino acids in P6 - P1' that vary with the species of potyvirus, e.g. Glu-Xaa-Xaa-Tyr-Xaa-Gln-/-(Ser or Gly) for the enzyme from tobacco etch virus. The natural substrate is the viral polyprotein, but other proteins and oligopeptides containing the appropriate consensus sequence are also cleaved.
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
139946-51-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
ClYVV
-
-
Manually annotated by BRENDA team
ClYVV
-
-
Manually annotated by BRENDA team
pepper vein banding potyvirus
-
-
-
Manually annotated by BRENDA team
PPV
-
-
Manually annotated by BRENDA team
potato virus Y PVY
PVY
-
-
Manually annotated by BRENDA team
turnip mosaic potyvirus
TuMV
-
-
Manually annotated by BRENDA team
turnip mosaic potyvirus TuMV
TuMV
-
-
Manually annotated by BRENDA team
polyprotein, part of the nuclear inclusion protein gene NIa of Vanilla mosaic virus-CI; VanMV, isolate from Vanilla tahitensis
SwissProt
Manually annotated by BRENDA team
ZYMV
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-Arg-Glu-Thr-Val-Arg-Phe-Gln-Ser-Asp-amide + H2O
?
show the reaction diagram
acetyl-Asn-Asn-Val-Arg-Phe-Gln-Ser-Leu-amide + H2O
acetyl-Asn-Asn-Val-Arg-Phe-Gln + Ser-Leu-amide
show the reaction diagram
acetyl-Glu-Pro-Thr-Val-Tyr-His-Gln-Thr-Leu-amide + H2O
?
show the reaction diagram
amyloid beta + H2O
?
show the reaction diagram
-
the enzyme cleaves monomeric and oligomeric amyloid beta at a similar rate in vitro
-
-
?
amyloid-beta + H2O
?
show the reaction diagram
-
-
-
-
?
amyloid-beta peptide + H2O
?
show the reaction diagram
-
degradation of oligomeric as well as monomeric forms of Amyloid-beta, presence of the consensus sequence, Val12-His-His-Gln15, near the presumptive alpha-secretase cleavage site of the amyloid-beta peptide
-
-
?
nuclear inclusion protein a + H2O
?
show the reaction diagram
P-sG polyprotein + H2O
?
show the reaction diagram
-
-
-
-
?
Pro-Thr-Thr-Glu-Leu-Tyr-Phe-Gln-Ser-Gly-Thr-Val-Asp-Arg-Arg + H2O
Pro-Thr-Thr-Glu-Leu-Tyr-Phe-Gln + Ser-Gly-Thr-Val-Asp-Arg-Arg
show the reaction diagram
RETVRFQSD + H2O
?
show the reaction diagram
-
-
-
?
TETVRFQSGTRR + H2O
?
show the reaction diagram
-
-
-
?
turnip mosaic virus polyprotein + H2O
?
show the reaction diagram
WDGGEVAHQAGESV + H2O
WDGGEVAHQ + AGESV
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
amyloid-beta + H2O
?
show the reaction diagram
-
-
-
-
?
nuclear inclusion protein a + H2O
?
show the reaction diagram
P-sG polyprotein + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NEM
-
does not completely inhibit NIa activity
additional information
-
no competitive inhibition of the proteolytic activity by the presence of excess of different protease mutants
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.217
acetyl-Arg-Glu-Thr-Val-Arg-Phe-Gln-Ser-Asp-amide
-
-
-
0.216
acetyl-Asn-Asn-Val-Arg-Phe-Gln-Ser-Leu-amide
-
-
-
0.51 - 2.83
acetyl-Glu-Pro-Thr-Val-Tyr-His-Gln-Thr-Leu-amide
0.069 - 0.271
Pro-Thr-Thr-Glu-Leu-Tyr-Phe-Gln-Ser-Gly-Thr-Val-Asp-Arg-Arg
0.082 - 0.108
TETVRFQSGTRR
0.05 - 11.4
WDGGEVAHQAGESV
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.079
acetyl-Arg-Glu-Thr-Val-Arg-Phe-Gln-Ser-Asp-amide
tobacco vein mottling virus
-
-
-
0.071
acetyl-Asn-Asn-Val-Arg-Phe-Gln-Ser-Leu-amide
tobacco vein mottling virus
-
-
-
0.12 - 1.01
acetyl-Glu-Pro-Thr-Val-Tyr-His-Gln-Thr-Leu-amide
0.033 - 0.18
Pro-Thr-Thr-Glu-Leu-Tyr-Phe-Gln-Ser-Gly-Thr-Val-Asp-Arg-Arg
0.092 - 0.094
TETVRFQSGTRR
0.002 - 0.23
WDGGEVAHQAGESV
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.87 - 1.12
TETVRFQSGTRR
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
amino acid and nucleotide identity percentages across the whole coat protein shown, the coat protein core region and the 3' non-translated region indicated, sequence identity to several Potyvirus strains determined, phylogenetic tree analysis performed
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
predominantly
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23000
-
mass spectrometry, N-terminal amino acid sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
three-dimensional structure of TVMV protease, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
-
the protease activity of NIa-Pro can be modulated by phosphorylation at Ser129
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant mutant TVMV protease K65A/K67A/C151A in complex with a canonical peptide substrate RETVRFQSD, mixing of protein solution with 10 mg/ml protein with a fivefold molar excess of the peptide substrate RETVRFQSD, crystallization from a solution consisting of 0.2 M potassium formate and 20% PEG 3350, space group P212121, X-ray diffraction structure determination and analysis at 1.7 A resolution, the 20-residue C-terminus of TVMV protease is disordered, molecular replacement, using the crystal structure of TEV protease, PDB code 1Q31
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15
turnip mosaic potyvirus
-
instable above
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
an expression cassette in which EGFP protein is expressed downstream of NIa protease and site F is transcribed within Escherichia coli strain BL21 (DE3)
-
cloning of NIa in Escherichia coli, overexpression of HA-tagged NIa in rat B103 neuroblastoma cells
-
co-expression of NIa-Pro and Vg-Pro in Escherichia coli
-
expressed in Escherichia coli BL21 pLysS cells
pepper vein banding potyvirus
-
expressed in HEK-293T cells and Mus musculus brain
-
expressed in Nicotiana benthamiana and Arabidopsis thaliana leaves
-
expressed in yeast
-
expression in Escherichia coli
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C151A
-
site-directed mutagenesis, a NIa-Pro active site mutant
E191A
-
site-directed mutagenesis, a VPg-Pro mutant
S129A
-
site-directed mutagenesis, a NIa-Pro phosphorylation-deficient mutant
S129D
-
site-directed mutagenesis, a NIa-Pro phosphorylation-mimic mnutant
W143A
-
site-directed mutagenesis, the NIa-Pro W143A mutant can bind the substrate with almost equal affinity as that of the wild-type protease, although its ability to catalyze the cleavage reaction is highly reduced
E189L
-
no self-cleavage
C151A
catalytically inactive mutant TVMV protease
K65A/K67A/C151A
catalytically inactive mutant TVMV protease
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture