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Information on EC 3.4.19.3 - pyroglutamyl-peptidase I and Organism(s) Pyrococcus furiosus and UniProt Accession O73944
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3.4.19.3 pyroglutamyl-peptidase ISpecify your search results
Word Map
- 3.4.19.3
- trh
- lymphocyte
- thymocytes
-
thyrotropin-releasing
- cd8
- thyrotropin
-
trh-degrading
- amyloliquefaciens
-
deblocked
- analysis
- thyroliberin
- his-pro
-
diketopiperazine
-
l-pyroglutamic
- phytolacca
- hermes
-
pancreatitis-associated
-
adipokinetic
- litoralis
-
post-proline
- protein-i
-
cyclo
-
3.4.21.26
- luliberin
-
intrathymic
- medicine
The taxonomic range for the selected organisms is: Pyrococcus furiosus
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Reaction Schemes
release of an N-terminal pyroglutamyl group from a polypeptide, the second amino acid generally not being Pro
Synonyms
pgp-1, pap-i, pyroglutamate aminopeptidase, pyroglutamyl aminopeptidase, pyroglutamyl peptidase i, pcp-0sh, pyrase, pyrrolidone carboxyl peptidase, pyrrolidonyl arylamidase, pyroglutamyl-peptidase i, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
5-oxoprolyl-peptidase
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-
-
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aminopeptidase, pyroglutamate
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-
-
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L-pyrrolidonecarboxylate peptidase
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-
-
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pyroglutamate aminopeptidase
pyroglutamidase
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-
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pyroglutamyl aminopeptidase
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-
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pyrrolidone-carboxylate peptidase
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-
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pyrrolidonecarboxy peptidase
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-
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pyrrolidonecarboxylyl peptidase
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-
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pyrrolidonyl peptidase
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pyroglutamate aminopeptidase
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
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-
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PATHWAY SOURCE
PATHWAYS
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CAS REGISTRY NUMBER
COMMENTARY
9075-21-2
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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
L-pyroglutamyl p-nitroanilide + H2O
L-pyroglutamate + p-nitroaniline
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-
-
-
?
L-pyroglutamyl-p-nitroanilide + H2O
L-pyroglutaminic acid + p-nitroaniline
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-
-
?
additional information
?
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PGAP performs enzymatic deblocking of pyroglutamylated immunoglobulins. Unfolding of immunoglobulins is required for excision of the pyroglutamate. The enzyme requires the substrate in an extended conformation, which is not fullfiled in the native form of immunoglobulins
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-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
methanol
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a methanol based deblocking solution preserves enzymatic activity, provides conditions compatible with sequencing and enhances deblocking of electroblotted samples
dithiothreitol
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strongly stimulates activity of wild-type enzyme and mutant enzyme C188S and shifts the pH-optimum to higher temperatures by about 10°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
75 - 105
75°C: about 60% of maximal activity, 105°C: about 90% of maximal activity
60 - 100
-
60°C: about 40% of maximal activity, 100°C: about 90% of maximal activity, wild-type enzyme, in presence of dithiothreitol
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
22936
x * 22936, ionspray triple quadrupole mass spectrometry, carboxymethylated enzyme
22940
ionspray triple quadrupole mass spectrometry, carboxymethylated enzyme
45643
x * 45643, ionspray triple quadrupole mass spectrometry, native enzyme
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
monomer
tetramer
?
x * 22936, ionspray triple quadrupole mass spectrometry, carboxymethylated enzyme
?
x * 45643, ionspray triple quadrupole mass spectrometry, native enzyme
dimer
-
the mutant enzyme C142S/C188S is monomeric below pH 2.7, dimeric around pH 3 and tetrameric above pH 4.5
dimer
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the mutant enzyme C142S/C188S is tetrameric above pH 4. The fraction of the dimeric form increases with increasing acidity below pH 4 and then the protein dissociates completely into a monomeric form at pH 2.5
monomer
-
the mutant enzyme C142S/C188S is monomeric below pH 2.7, dimeric around pH 3 and tetrameric above pH 4.5
monomer
-
the mutant enzyme C142S/C188S is tetrameric above pH 4. The fraction of the dimeric form increases with increasing acidity below pH 4 and then the protein dissociates completely into a monomeric form at pH 2.5
tetramer
-
the mutant enzyme C142S/C188S is monomeric below pH 2.7, dimeric around pH 3 and tetrameric above pH 4.5
tetramer
-
the mutant enzyme C142S/C188S is tetrameric above pH 4. The fraction of the dimeric form increases with increasing acidity below pH 4 and then the protein dissociates completely into a monomeric form at pH 2.5
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PCP-0SH polar mutants C142S/C188S/E192D andC142S/C188S/E192Q crystallize at a 6.5% PEG4000, while the apolar mutants C142S/C188S/E192A, C142S/C188S/E192I and C142S/C188S/E192V crystallize at a 5.7-6.0% PEG4000. The protein molecules crystallize in two different space groups. E192Q and E192V form isomorphic monoclinic crystals in the space group P2(1), which agree with those of the wild-type PCP and cysteine-free PCP-0SH (C142S/C188S), while E192A, E192D, and E192I form orthorhombic crystals in the space group P2(1)2(1)2(1). In both crystal systems, four subunit (monomer) molecules are contained in the asymmetric unit. A systematic analysis of individual structures indicates that the mutation does not have any significant effect on the overall structure
crystal structure of wild-type and mutant enzyme C142S/C188S determined at 2.2 and 2.7 A resolution
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A199P
alpha6-helix region of A199P in the D1 state (initial denatured state) is partially unprotected, while some hydrophobic residues are protected against H/D exchange, although these hydrophobic residues are unprotected in the wild-type protein. Structure of A199P in the D1 state forms a temporary stable denatured structure with a non-native hydrophobic cluster and the unstructured alpha6-helix
C142S/C188S/E192A
at acidic pH the mutant enzyme is less stable than cysteine-free mutant C142S/C188S. At alkaline pH the mutant enzyme is more stable than cysteine-free mutant C142S/C188S. The thermal stability of the mutant enzyme at pH 2.15, pH 3.04 and pH 7.3 is less than that of the cysteine-free mutant enzyme C142S/C188S. At pH 8.7 and 9.6 the thermal stability of mutant enzyme is higher than that of the cysteine-free mutant C142S/C188S
C142S/C188S/E192D
at acidic pH the mutant enzyme is less stable than cysteine-free mutant C142S/C188S. The thermal stability of the mutant enzyme at pH 2.15, pH 3.04, pH 7.3, pH 8.7 and pH 9.6 is less than that of the cysteine-free mutant enzyme C142S/C188S
C142S/C188S/E192I
at acidic pH the mutant enzyme is less stable than cysteine-free mutant C142S/C188S. At alkaline pH the mutant enzyme is more stable than cysteine-free mutant C142S/C188S. The thermal stability of the mutant enzyme at pH 2.15, pH 3.04 and pH 7.3 is less than that of the cysteine-free mutant enzyme C142S/C188S. At pH 8.7 and 9.6 the thermal stability of mutant enzyme is higher than that of the cysteine-free mutant C142S/C188S
C142S/C188S/E192Q
at acidic pH the mutant enzyme is less stable than cysteine-free mutant C142S/C188S. At alkaline pH the mutant enzyme is more stable than cysteine-free mutant C142S/C188S. The thermal stability of the mutant enzyme at pH 2.15, pH 3.04 and pH 7.3 is less than that of the cysteine-free mutant enzyme C142S/C188S. At pH 8.7 and 9.6 the thermal stability of mutant enzyme is higher than that of the cysteine-free mutant C142S/C188S
C142S/C188S/E192V
at acidic pH the mutant enzyme is less stable than cysteine-free mutant C142S/C188S. At alkaline pH the mutant enzyme is more stable than cysteine-free mutant C142S/C188S. The thermal stability of the mutant enzyme at pH 2.15, pH 3.04 and pH 7.3 is less than that of the cysteine-free mutant enzyme C142S/C188S. At pH 8.7 and 9.6 the thermal stability of mutant enzyme is higher than that of the cysteine-free mutant C142S/C188S
Cys144Ser/Cys188Ser
cysteine-free variant. The 114-208 segment of the mutant folds into a stable compact structure with non-native helix-helix association in the D1 state. In the folding process from the D1 state to the native state, the alpha4- and alpha6-helices become separated and the central beta-sheet is folded between these helices. The non-native interaction between the alpha4- and alpha6-helices may be responsible for the unusually slow folding of the mutant
C142S/C188S
C188S
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activity is reduced by one-fourth relative to the activity of the wild-type enzyme
C142S/C188S
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the mutant enzyme is used in unfolding and refolding experiments to exclude the complexity in reaction due to oxidation of SH groups
C142S/C188S
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small thermodynamic stability of the mutant enzyme C142S/C188S at low pH. The mutant enzyme is monomeric below pH 2.7, dimeric around pH 3 and tetrameric above pH 4.5. The heat-denaturation of the mutant enzyme is completely reversible at pH 2.3, although the unfolding-refolding reactions are characterized by extremely slow kinetics
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10.5
at alkaline pH the mutant enzyme enzymes C142S/C188S/E192A, C142S/C188S/E192I, C142S/C188S/E192V and C142S/C188S/E192Q are more stable than cysteine-free mutant C142S/C188S
667733
2.4
at acidic pH the mutant enzymes C142S/C188S/E192A, C142S/C188S/E192I, C142S/C188S/E192V, C142S/C188S/E192D and C142S/C188S/E192Q are less stable than cysteine-free mutant C142S/C188S
667733
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
102
peak temperature on the differential scanning calorimetry is 101.7°C, pH 9,6, cysteine-free mutant enzyme C142S/C188S
59
peak temperature on the differential scanning calorimetry is 59.3°C, pH 2.15, cysteine-free mutant enzyme C142S/C188S
79
peak temperature on the differential scanning calorimetry is 78.9°C, pH 3.04 cysteine-free mutant enzyme C142S/C188S
89
47
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thermodynamics of heat denaturation of the monomeric enzyme form of mutant enzyme C142S/C188S at pH 2.3. The mechanism of refolding is a two-state process. The equilibrium establishes with a relaxation time of 5080 s at Tm = 46.5°C
additional information
89
peak temperature on the differential scanning calorimetry is 77.6°C, pH 7.3, cysteine-free mutant enzyme C142S/C188S
89
peak temperature on the differential scanning calorimetry is 88.8°C, pH 8,7, cysteine-free mutant enzyme C142S/C188S
additional information
the thermal stability of the mutant enzymes C142S/C188S/E192A, C142S/C188S/E192I, C142S/C188S/E192V, C142S/C188S/E192D and C142S/C188S/E192Q at pH 2.15, pH 3.04 and pH 7.3 is less than that of the cysteine-free mutant enzyme C142S/C188S. At pH 8.7 and 9.6 the thermal stability of mutant enzymes C142S/C188S/E192A, C142S/C188S/E192I, C142S/C188S/E192V and C142S/C188S/E192Q is higher than that of the cysteine-free mutant C142S/C188S. The thermal stability of mutant enzyme C142S/C188S/E192D at pH 8.7 and 9.6 is less than that of cysteine-free mutant enzyme C142S/C188S
additional information
-
the thermal stability of the mutant enzymes C142S/C188S/E192A, C142S/C188S/E192I, C142S/C188S/E192V, C142S/C188S/E192D and C142S/C188S/E192Q at pH 2.15, pH 3.04 and pH 7.3 is less than that of the cysteine-free mutant enzyme C142S/C188S. At pH 8.7 and 9.6 the thermal stability of mutant enzymes C142S/C188S/E192A, C142S/C188S/E192I, C142S/C188S/E192V and C142S/C188S/E192Q is higher than that of the cysteine-free mutant C142S/C188S. The thermal stability of mutant enzyme C142S/C188S/E192D at pH 8.7 and 9.6 is less than that of cysteine-free mutant enzyme C142S/C188S
additional information
alpha6-helix (from Ser188 to Glu205) of PCP-0SH is stably formed in the D1 state
additional information
-
alpha6-helix (from Ser188 to Glu205) of PCP-0SH is stably formed in the D1 state
additional information
-
the heat denaturation of wild-type enzyme and mutant enzymes C188S and C142S/C188S is highly reversible in the dimeric forms, but completely irreversible in the tetrameric form
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
subunit interactions play an important role in stabilizing the enzyme in addition to the intrinsic enhanced stability of its monomer
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wild-type and mutant enzyme C142S/C188S, the anomalous high stability of the hyperthermophilic enzyme originates from the unusually slow rate of unfolding reaction during treatment with guanidine-hydrochloride
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
guanidine-HCl
approximately 70% of the original activity is retained after preincubation with 10 mM DTT, 50 mM Na-phosphate buffer (pH 7.0) containing less than either 0.01% SDS, 1 M urea, or 1 M guanidine-HCl at 37°C for 15 min
SDS
approximately 70% of the original activity is retained after preincubation with 10 mM DTT, 50 mM Na-phosphate buffer (pH 7.0) containing less than either 0.01% SDS, 1 M urea, or 1 M guanidine-HCl at 37°C for 15 min
urea
approximately 70% of the original activity is retained after preincubation with 10 mM DTT, 50 mM Na-phosphate buffer (pH 7.0) containing less than either 0.01% SDS, 1 M urea, or 1 M guanidine-HCl at 37°C for 15 min
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
PGAP expressed from a pET3a-series vector in Escherichia coli BL21(DE) cells
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
the heat denaturation of wild-type enzyme and mutant enzymes C188S and C142S/C188S is highly reversible in the dimeric forms, but completely irreversible in the tetrameric form
-
the heat-denaturation of the mutant enzyme C142S/C188S is completely reversible at pH 2.3, although the unfolding-refolding reactions are characterized by extremely slow kinetics
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
C-terminal alpha-helix in the D1 state plays an important role in retaining the D1 state under the stable conditions and in correctly folding into the native structure of PCP-0SH
additional information
-
C-terminal alpha-helix in the D1 state plays an important role in retaining the D1 state under the stable conditions and in correctly folding into the native structure of PCP-0SH
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ogasahara, K.; Nakamura, M.; Nakura, S.; Tsunasawa, S.; Kato, I.; Yoshimoto, T.; Yutani, K.
The unusually slow unfolding rate causes the high stability of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyroccoccus furiosus: equilibrium and kinetic studies of guanidine hydrochloride-induced unfolding and refolding
Biochemistry
37
17537-17544
1998
Bacillus amyloliquefaciens, Pyrococcus furiosus
Ogasahara, K.; Khechinashvili, N.N.; Nakamura, M.; Yoshimoto, T.; Yutani, K.
Thermal stability of pyrrolidone carboxyl peptidases from the hyperthermophilic archaeon, Pyrococcus furiosus
Eur. J. Biochem.
268
3233-3242
2001
Pyrococcus furiosus
Tanaka, H.; Chinami, M.; Mizushima, T.; Ogasahara, K.; Ota, M.; Tsukihara, T.; Yutani, K.
X-ray crystalline structures of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus, and its Cys-free mutant
J. Biochem.
130
107-118
2001
Pyrococcus furiosus
Kaushik, J.K.; Ogasahara, K.; Yutani, K.
The unusually slow relaxation kinetics of the folding-unfolding of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus
J. Mol. Biol.
316
991-1003
2002
Pyrococcus furiosus
Kaushik, J.K.; Iimura, S.; Ogasahara, K.; Yamagata, Y.; Segawa, S.; Yutani, K.
Completely buried, non-ion-paired glutamic acid contributes favorably to the conformational stability of pyrrolidone carboxyl peptidases from hyperthermophiles
Biochemistry
45
7100-7112
2006
Pyrococcus furiosus (O73944), Pyrococcus furiosus
Iimura, S.; Umezaki, T.; Takeuchi, M.; Mizuguchi, M.; Yagi, H.; Ogasahara, K.; Akutsu, H.; Noda, Y.; Segawa, S.; Yutani, K.
Characterization of the denatured structure of pyrrolidone carboxyl peptidase from a hyperthermophile under nondenaturing conditions: role of the C-terminal alpha-helix of the protein in folding and stability
Biochemistry
46
3664-3672
2007
Pyrococcus furiosus (O73944), Pyrococcus furiosus
Hellstroem, J.L.; Vehniaeinen, M.; Mustonen, M.; Loevgren, T.; Lamminmaeki, U.; Hellman, J.
Unfolding of the immunoglobulin light and heavy chains is required for the enzymatic removal of N-terminal pyroglutamyl residues
Biochim. Biophys. Acta
1764
1735-1740
2006
Pyrococcus furiosus
Tsunasawa, S.; Nakura, S.; Tanigawa, T.; Kato, I.
Pyrrolidone carboxyl peptidase from the hyperthermophilic archaeon Pyrococcus furiosus: cloning and overexpression in Escherichia coli of the gene, and its application to protein sequence analysis
J. Biochem.
124
778-783
1998
Pyrococcus furiosus (O73944), Pyrococcus furiosus
Mizuguchi, M.; Takeuchi, M.; Ohki, S.; Nabeshima, Y.; Kouno, T.; Aizawa, T.; Demura, M.; Kawano, K.; Yutani, K.
Structural characterization of a trapped folding intermediate of pyrrolidone carboxyl peptidase from a hyperthermophile
Biochemistry
51
6089-6096
2012
Pyrococcus furiosus (O73944)
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