Information on EC 3.4.22.68 - Ulp1 peptidase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY hide
3.4.22.68
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RECOMMENDED NAME
GeneOntology No.
Ulp1 peptidase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of the alpha-linked peptide bond in the sequence Gly-Gly-/-Ala-Thr-Tyr at the C-terminal end of the small ubiquitin-like modifier (SUMO) propeptide, Smt3, leading to the mature form of the protein. A second reaction involves the cleavage of an epsilon-linked peptide bond between the C-terminal glycine of the mature SUMO and the lysine epsilon-amino group of the target protein
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY hide
252852-50-9
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
His6-Smt3-hemagglutinin fusion protein + H2O
?
show the reaction diagram
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?
His6-Smt3-Leu-beta-galactosidase + H2O
?
show the reaction diagram
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-
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?
His6-Smt3-Met-beta-galactosidase + H2O
?
show the reaction diagram
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?
small ubiquitin-like modifier protein + H2O
?
show the reaction diagram
small ubiquitin-related modifier-CENP-I + H2O
small ubiquitin-related modifier-protein + CENP-I
show the reaction diagram
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SUMO-specific proteases, SENPs, reversibly remove small ubiquitin-related modifier-protein, SUMO, from the SUMOylated proteins
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r
small ubiquitin-related modifier-Mdm2 + H2O
small ubiquitin-related modifier-protein + Mdm2
show the reaction diagram
small ubiquitin-related modifier-protein + H2O
small ubiquitin-related modifier-protein + protein
show the reaction diagram
SMT3precursor + H2O
?
show the reaction diagram
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?
SUMO-1 protein + H2O
?
show the reaction diagram
SUMO-GFP fusion substrate + H2O
SUMO + GFP
show the reaction diagram
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pH 8.0, 25°C, in the presence of 5 mM 2-mercaptoethanol
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?
SUMO-MMP13 + H2O
?
show the reaction diagram
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cleavage occurs only to 60%
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
small ubiquitin-related modifier-CENP-I + H2O
small ubiquitin-related modifier-protein + CENP-I
show the reaction diagram
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SUMO-specific proteases, SENPs, reversibly remove small ubiquitin-related modifier-protein, SUMO, from the SUMOylated proteins
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r
small ubiquitin-related modifier-Mdm2 + H2O
small ubiquitin-related modifier-protein + Mdm2
show the reaction diagram
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co-localization of SENP2 with SUMO conjugated Mdm2. SUMO conjugation of Mdm2 induces its co-localization and association with SENP2 at the PML bodies
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r
small ubiquitin-related modifier-protein + H2O
small ubiquitin-related modifier-protein + protein
show the reaction diagram
SMT3precursor + H2O
?
show the reaction diagram
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?
SUMO-1 protein + H2O
?
show the reaction diagram
additional information
?
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the enzyme is specifically required for cell cycle progression
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-[(4-[[2-([(2R,3R)-3-[benzyl(cyclohexa-1,3-dien-1-ylmethyl)carbamoyl]-3-chlorooxiran-2-yl]carbonyl)-2-(carboxymethyl)hydrazinyl]carbonyl]benzyl)carbamoyl]benzoic acid
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partial inhibition of the enzyme
3-[(4-[[2-[(2E)-4-[bis(naphthalen-1-ylmethyl)amino]-4-oxobut-2-enoyl]-2-(carboxymethyl)hydrazinyl]carbonyl]benzyl)carbamoyl]benzoic acid
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3-[(4-[[2-[4-[bis(naphthalen-1-ylmethyl)amino]-2,3-dichloro-4-oxobutanoyl]-2-(carboxymethyl)hydrazinyl]carbonyl]benzyl)carbamoyl]benzoic acid
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the chlorohydrin form of JCP-666 may inhibit the target SENP by SN2-like displacement of the chloride by the active site cysteine
3-[(4-[[2-[4-[bis(naphthalen-1-ylmethyl)amino]-3-chloro-2-hydroxy-4-oxobutanoyl]-2-(carboxymethyl)hydrazinyl]carbonyl]benzyl)carbamoyl]benzoic acid
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i.e. JCP-666
Gu-HCl
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500 mM reduces cleavage to 60%, 1 M reduces cleavage to 0%
hSUMO-VS
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human SUMO protein modified with a vinyl sulfone reactive group after the C-terminal di-glycine, contains the full length SUMO protein fused to a reactive vinyl sulfone group, an irreversible inhibitor of SENP proteases
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N-ethylmaleimide
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NEM, blocks SENP activity by acting as a general alkylating agent that modifies the active site cysteine in parasite lysates
NaCl
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500 mM reduces cleavage to 60%, 1 M reduces cleavage to 30%
Ubiquitin aldehyde
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Urea
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2 M reduces cleavage to 95%, 3 M reduces cleavage to 5%
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
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Michaelis-Menten steady-state kinetics are performed for SENP6 by introduction of S9C and C52A point mutants into SUMO1 and SUMO1 mutant A68N/H71D to allow for fluorophore addition
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
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assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
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parasites in human red blood cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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the NH2-terminal domain of the enzyme, residues between position 144 and 346, includes sequences necessary and sufficient to concentrate Ulp1 at nuclear envelope
Manually annotated by BRENDA team
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SENP6 and SENP7
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 31000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ulp1-Smt3 crystal structure, hanging drop vapor diffusion
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, 50% glycerol, 75 mM Tris-HCl pH 8.0, 0.5 mM DTT, 1 mM EDTA, 6 months, no detectable decay of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant catalytic domains of human SENP2-(364-589), SENP6-(637-1112), and SENP7-(662-984) and of mutant SENP-7s from Escherichia coli by metal affinity chromatography and gel filtration
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recombinant FLAG-tagged and maltose-binding protein-fused isozyme Ulp1, containing a TEV protease recognition site, from Escherichia coli strain BL21 by affinity chromatography
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recombinant His-tagged SUMO-S-GroQ by nickel affinity chromatography to over 95% purity, SUMO is clevaed off by co-expressed SUMO-fused SUMO-specific protease Ulp1
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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expression of catalytic domains of human SENP2-(364-589), SENP6-(637-1112), and SENP7-(662-984) and of mutant SENP-7s in Escherichia coli
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expression of FLAG-SMT3IP1 in COS-7 cells, coexpression with His6-ubiquitin. Overexpression of SMT3IP1 in cells results in the accumulation of Mdm2 in the nucleolus and increased stability of the p53 protein
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expression of SENP1 and SENP2 isozymes in HCT-116 cells
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expression of SUMO-S-GroQ fusion protein, i.e. SUMO fused to the Q domain of Drosophila melanogaster Groucho, containing sequences encoding the mature form of SUMO followed by a 6-His tag and a multiple cloning site harbouring the fused protein S-GroQ, co-expression of SUMO-fused SUMO-specific protease Ulp1
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expression of the wild-type full-length PfSENP1 and the catalytic domain of PfSENP1
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expression of wild-type and mutant enzymes, expression of the SBS domain as a SBS-GFP fusion protein
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gene ESD4, DNA and amino acid sequence determination, genotyping; gene ULP1A, DNA and amino acid sequence determination, genotyping, recombinant expression in Arabidopsis thaliana plants and in leaves of Nicotioana bentamiana using transfection via Agrobacterium tumefaciens. An expressed GFP-ELS1 construct has a generally non-nuclear localization, being detectable at membranes and in the cytoplasm
isozyme Ulp1, expression of recombinant FLAG-tagged and maltosse-binding protein-fused isozyme Ulp1, containing a TEV protease recognition site, in Escherichia coli strain BL21, recombinant expression of GST-tagged isozyme Ulp1
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C461S
site-directed mutagenesis, ELS1C461S is not capable of cleaving the extension oV the carboxyl terminus of SUMO1
K691A
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site-directed mutagenesis of SENP7
K691E
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site-directed mutagenesis of SENP7
P686G/P687G/P688G/P689G
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site-directed mutagenesis of SENP7
C580S
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site-directed mutagenesis, generation of a catalytically inactive mutant of Ulp1, the mutant is greatly enriched at the septin ring of dividing yeast cells. The 218-amino acid, substrate-trapping mutant of the catalytic domain of Ulp1 is necessary and sufficient for septin localization
D451N
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the mutation destroys an essential salt bridge formed between Smt3 and Ulp1
D451N/C580S
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site-directed mutagenesis, abolished accumulation of the full-length Ulp1 double-mutant at the septin ring
I435V/N450S/I504T/C580S
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site-directed mutagenesis, the mutant shows a reduced ability to enrich at the septin ring
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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substrate-trapping Ulp1(3)(C580S) interacts robustly with human SUMO1, SUMO2 and SUMO2 chains, making it a potentially useful tool for the analysis and purification of SUMO-modified proteins
medicine
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there may be a connection between a defect in SUMO-1 conjugation to the PML protein and acute promyelocytic leukemia (ALP). Specific Ulp inhibitors can therefore have therapeutic value for ALP
molecular biology