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Information on EC 3.4.22.68 - Ulp1 peptidase and Organism(s) Saccharomyces cerevisiae and UniProt Accession Q02724
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3.4.22.68 Ulp1 peptidaseSpecify your search results
Word Map
- 3.4.22.68
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desumoylation
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ubiquitin-related
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senps
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deconjugation
- sumo1
- isopeptidase
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sentrin-specific
- hypoxia
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ni-nta
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sumo-conjugating
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sumo2/3
- piasy
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sumo-dependent
- nemo
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sumo-modified
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sumoylation-deficient
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sumo-targeted
- synthesis
- medicine
- biotechnology
- molecular biology
The taxonomic range for the selected organisms is: Saccharomyces cerevisiae
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
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
Synonyms
senp1, senp2, sumo protease, senp6, sumo-specific protease, senp7, smt3ip1, sumo protease 1, ulp1 protease, sumo-specific protease 2, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
C48.001
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Small ubiquitin-related modifier protein 1 conjugate proteinase
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Smt3-protein conjugate proteinase
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Ulp1
Ulp1 endopeptidase
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Ulp1
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY
252852-50-9
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
SUMO-GFP fusion substrate + H2O
SUMO + GFP
pH 8.0, 25°C, in the presence of 5 mM 2-mercaptoethanol
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?
small ubiquitin-like modifier protein + H2O
?
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Ulp1 catalyzes the proteolytic processing of SUMO to its mature form
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?
small ubiquitin-like modifier protein + H2O
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Ulp1 catalyzes the proteolytic processing of SUMO to its mature form
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?
SUMO-1 protein + H2O
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Ulp1 catalyzes two essential functions in the SUMO pathway: 1. the processing of full-length SUMO to its mature form and 2. deconjugation of SUMO from target proteins. Ulp1 can proteolyze large folded SUMO-conjugated proteins without altering their structure
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?
SUMO-1 protein + H2O
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the enzyme plays an essential role in the G2/M phase of the cell cycle
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?
additional information
?
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Ulp1 liberates poly-Smt3 from a substrate chain. In vitro, Ulp1 is highly active even in very low concentrations. Substrate specificity analysis of immobilized recombinant enzyme, overview
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?
additional information
?
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the NH2-terminal regulatory domain of Ulp1 restricts Ulp1 activity towards certain sumoylated proteins while enabling the cleavage of others, the COOH-terminal catalytic domain of Ulp1 is both necessary and sufficient for the essential function of the protein in cell cycle progression and for Smt3 precursor cleavage
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?
additional information
?
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the enzyme is specifically required for cell cycle progression
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?
additional information
?
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Ulp1 is responsible for both removing SUMO/Smt3 from specific target proteins and for processing precursor SUMO into its conjugation-competent form. Multiple features in the catalytic domain of Ulp1 affect SUMO interactions, analysis of features of Ulp1 required for substrate targeting, structure-function analysis, overview. D451 is required for targeting of sumoylated proteins and the C580S mutation is required for retention of Ulp1 at the septin ring. Kap121-independent SUMO-targeting information resides in the catalytic domain of Ulp1. The Ulp1 Kap121-interacting domain (region 1), the Ulp1 Kap60/Kap95-interacting domain (region 2) and the catalytic domain (region 3) fail to interact with the Smt3-binding domain
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?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
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the enzyme is specifically required for cell cycle progression
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-
?
SUMO-1 protein + H2O
?
Ulp1 catalyzes two essential functions in the SUMO pathway: 1. the processing of full-length SUMO to its mature form and 2. deconjugation of SUMO from target proteins. Ulp1 can proteolyze large folded SUMO-conjugated proteins without altering their structure
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?
SUMO-1 protein + H2O
?
the enzyme plays an essential role in the G2/M phase of the cell cycle
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?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Gu-HCl
500 mM reduces cleavage to 60%, 1 M reduces cleavage to 0%
NaCl
500 mM reduces cleavage to 60%, 1 M reduces cleavage to 30%
additional information
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
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additional information
no inhibitory effects are observed with Triton X100 (1 M), imidazole (300 mM), reduced glutathione (20 mM), maltose (20 mM), glycerol (20% v/v), ethylene glycol (20% v/v), sucrose (20% w/v), ethanol (10% v/v)
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additional information
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no inhibitory effects are observed with Triton X100 (1 M), imidazole (300 mM), reduced glutathione (20 mM), maltose (20 mM), glycerol (20% v/v), ethylene glycol (20% v/v), sucrose (20% w/v), ethanol (10% v/v)
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
constructed strain contains a single ulp1-I615N mutation
SwissProt
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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Ulp1 localizes predominantly to nuclear pore complexes but also deconjugates sumoylated septins at the bud-neck of dividing cells. A 218-amino acid, substrate-trapping mutant of the catalytic domain of Ulp1, Ulp1(3) C580S is necessary and sufficient for septin localization
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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
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Ulp1 localizes predominantly to nuclear pore complexes but also deconjugates sumoylated septins at the bud-neck of dividing cells
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
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an ulp1 temperature-sensitive strain ulp1-333SGG with inactivated enzyme Ulp1 at restrictive temperature accumulates unprocessed SUMO protein
physiological function
physiological function
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Ulp1 facilitates sumoylation by processing precursor SUMO into its conjugation competent form. Conversely, Ulp1 also facilitates desumoylation by removing SUMO from nuclear and cytosolic proteins after conjugation. The essential small ubiquitin-like modifier, SUMO, protease Ulp1 is responsible for both removing SUMO/Smt3 from specific target proteins and for processing precursor SUMO into its conjugation-competent form
physiological function
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the enzyme Ulp1 does not appear to play a major role in the inactivation of the SUMO stress response consisting of consist of a simple SUMO conjugation-deconjugation cycle, overview. Isozyme Ulp1 is required for activation of the SUMO pro-protein, but SUMO stress response does not require proteasome-mediated degradation or Ulp1 activity
physiological function
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the catalytic UD domains of both Saccharomyces cerevisiae ScUlp1 and Kluyveromyces marxianus KmUlp1 show a high degree of sequence conservation, complement a Ulp1 deletion mutant in vivo, and process a SUMO precursor in vitro. Catalytically inactive recombinant fragments of the UD domains are able to efficiently bind a variety of purified SUMO isoforms and bind immobilized SUMO1 with nanomolar affinity
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
additional information
immobilization of isozyme Ulp1 via its C-terminal FLAG tag
additional information
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generation of a collection of GFP-tagged Ulp1 truncations and domains that were expressed under control of the Ulp1 promoter. truncations and domains of Ulp1, that retain substrate targeting information, also localize to the septin ring in G2/M-arrested cells. Usage of the targeting and SUMO-binding properties of Ulp1(3)(C580S) to purify Smt3-modified proteins from cell extracts. Deletion of the entire SBS domain on the localization of Ulp1(3)(C580S). The Ulp1(3)(C580S)SBSDELTA construct does not localize to the septin ring in the majority of cells
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
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant FLAG-tagged and maltose-binding protein-fused isozyme Ulp1, containing a TEV protease recognition site, from Escherichia coli strain BL21 by affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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
expression of wild-type and mutant enzymes, expression of the SBS domain as a SBS-GFP fusion protein
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
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
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
molecular biology
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usage of the targeting and small ubiquitin-like modifier, SUMO, binding properties of Ulp1(3)(C580S) to purify Smt3-modified proteins from cell extracts
synthesis
synthesis
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comparison of Ulp1 protease in active upon expression as inclusion bodies and soluble protein. Fusion of the N-terminal selfassembling peptide GFIL8 to Ulp1 increases production of active inclusion bodies in Escherichia coli. Attachment of the N-terminal cellulose-binding module facilitates the immobilization on regenerated amorphous cellulose with a binding capacity up to about 235 mg protein per gram of cellulose. The immobilized soluble Ulp1 maintains about 42% initial cleavage activity with repetitive use, whereas the aggregated Ulp1 loses its cleavage capacity after cleaving the protein substrate once. Crosslinking of inclusion bodies using glutaraldehyde inactivates Ulp1
synthesis
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immobilization of Ulp1 as a tool for cleavage of the SUMO tag of recombinant proteins. Ulp1 immobilized on N-hydroxysuccinimide-activated Sepharose maintains 95% substrate-cleavage ability and significantly enhanced pH and thermal stability. The immobilized Ulp1 can tolerate 15% (v/v) DMSO and 20% (v/v) ethanol. It can be reused for more than 15 batch reactions with 90% activity retention
synthesis
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optimization of expression of the catalytic domain. Optimization of cultivation conditions at shake flask results in Ulp1 expression of 195 mg/l in TB medium. Ni-NTA affinity purification of Ulp1 using 0.1% Triton X-100, 0.01mM DTT, 0.02mM EDTA and 1% glycerol leads to a purity of about 95% with a recovery yield of 80% and specific activity of 398600 U/mg. The protease cleaves the SUMO tag even at 1:10,000 enzyme to substrate ratio The in vivo cleavage of SUMO tag via coexpression strategy also results in more than 80% cleavage of SUMO fusion protein
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Li, S.J.; Hochstrasser, M.
A new protease required for cell-cycle progression in yeast
Nature
398
246-251
1999
Saccharomyces cerevisiae (Q02724)
Mossessova, E.; Lima, C.D.
Ulp1-SUMO crystal structure and genetic analysis reveal conserved interactions and a regulatory element essential for cell growth in yeast
Mol. Cell
5
865-876
2000
Saccharomyces cerevisiae (Q02724), Saccharomyces cerevisiae
Li, S.J.; Hochstrasser, M.
The Ulp1 SUMO isopeptidase: distinct domains required for viability, nuclear envelope localization, and substrate specificity
J. Cell. Biol.
160
1069-1081
2003
Saccharomyces cerevisiae
Lima, C.D.
Ulp1 endopeptidase
Handbook of Proteolytic Enzymes (Barrett, A. J. ; Rawlings, N. D. ; Woessner, eds. )
2
1340-1344
2004
Schizosaccharomyces pombe (O42957), Saccharomyces cerevisiae (Q02724)
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Malakhov, M.P.; Mattern, M.R.; Malakhova, O.A.; Drinker, M.; Weeks, S.D.; Butt, T.R.
SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins
J. Struct. Funct. Genomics
5
75-86
2004
Saccharomyces cerevisiae (Q02724), Saccharomyces cerevisiae
Soustelle, C.; Vernis, L.; Freon, K.; Reynaud-Angelin, A.; Chanet, R.; Fabre, F.; Heude, M.
A new Saccharomyces cerevisiae strain with a mutant Smt3-deconjugating Ulp1 protein is affected in DNA replication and requires Srs2 and homologous recombination for its viability
Mol. Cell. Biol.
24
5130-5143
2004
Saccharomyces cerevisiae (Q02724)
Dobson, M.J.; Pickett, A.J.; Velmurugan, S.; Pinder, J.B.; Barrett, L.A.; Jayaram, M.; Chew, J.S.
The 2 microm plasmid causes cell death in Saccharomyces cerevisiae with a mutation in Ulp1 protease
Mol. Cell. Biol.
25
4299-4310
2005
Saccharomyces cerevisiae (Q02724), Saccharomyces cerevisiae
Elmore, Z.C.; Donaher, M.; Matson, B.C.; Murphy, H.; Westerbeck, J.W.; Kerscher, O.
Sumo-dependent substrate targeting of the SUMO protease Ulp1
BMC Biol.
9
74
2011
Saccharomyces cerevisiae
Eckhoff, J.; Dohmen, R.J.
In vitro studies reveal a sequential mode of chain processing by the yeast SUMO-specific protease Ulp2
J. Biol. Chem.
19
12268-12281
2015
Saccharomyces cerevisiae (Q02724)
Lewicki, M.C.; Srikumar, T.; Johnson, E.; Raught, B.
The S. cerevisiae SUMO stress response is a conjugation-deconjugation cycle that targets the transcription machinery
J. Proteomics
118
39-48
2015
Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4741/2
Liang, Q.; Huang, Z.; Zhang, Y.; Li, H.
Immobilization of Ulp1 protease on NHS-activated Sepharose a useful tool for cleavage of the SUMO tag of recombinant proteins
Biotechnol. Lett.
39
1025-1031
2017
Saccharomyces cerevisiae
Babbal, B.; Adivitiya, B.; Mohanty, S.; Khasa, Y.P.
Bioprocess optimization for the overproduction of catalytic domain of ubiquitin-like protease 1 (Ulp1) from S. cerevisiae in E. coli fed-batch culture
Enzyme Microb. Technol.
120
98-109
2019
Saccharomyces cerevisiae
Jiang, L.; Xiao, W.; Zhou, X.; Wang, W.; Fan, J.
Comparative study of the insoluble and soluble Ulp1 protease constructs as carrier free and dependent protein immobilizates
J. Biosci. Bioeng.
127
23-29
2019
Saccharomyces cerevisiae
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