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Information on EC 3.4.11.B4 - Pyrococcus horikoshii deblocking aminopeptidase

for references in articles please use BRENDA:EC3.4.11.B4
preliminary BRENDA-supplied EC number
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UNIPROT: O58255
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Word Map
The expected taxonomic range for this enzyme is: Pyrococcus horikoshii
Reaction Schemes
The enzyme exhibits aminopeptidase activity and with lower efficiency deblocking activity (i.e. hydrolysis of acetylated amino acids from the N-terminus of peptides). The turnover number with Ac-Ala-Ala-Ala is 260-540fold lower compared to the activity with Ala-Ala-Ala. The turnover number with Ac-Ala-Ala is 350fold lower compared to the activity with Ala-Ala. Low hydrolytic activity for peptide substrates longer than 10 residues. The enzyme is activated by Co2+ or Zn2+. Contains 1.32 mol of Ca2+ per mol of monomer.
Synonyms
deblocking aminopeptidase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
deblocking aminopeptidase
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Ala-Ala + H2O
L-alanine + L-alanine
show the reaction diagram
-
-
-
?
Ala-Ala-Ala + H2O
Ala-Ala + L-alanine
show the reaction diagram
Ala-Ala-Ala-Ala + H2O
Ala-Ala-Ala + L-alanine
show the reaction diagram
-
-
-
?
Ala-Ala-Ala-Ala-Ala + H2O
Ala-Ala-Ala-Ala + L-alanine
show the reaction diagram
-
-
-
?
N-acetyl-Ala-Ala-Ala + H2O
Ala-Ala + N-acetyl-L-alanine
show the reaction diagram
the activity (turnover number) with N-acetyl-Ala-Ala-Ala is 260fold lower compared to the activity with Ala-Ala-Ala
-
-
?
N-acetyl-L-alanyl 4-nitroanilide + H2O
N-acetyl-L-alanine + 4-nitroaniline
show the reaction diagram
-
-
-
?
N-acetyl-L-leucyl 4-nitroanilide + H2O
N-acetyl-L-leucine + 4-nitroaniline
show the reaction diagram
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
39.9
Ala-Ala
pH 7.5, 85°C
3.4 - 18.2
Ala-Ala-Ala
13.2
Ala-Ala-Ala-Ala
pH 7.5, 85°C
10.8
Ala-Ala-Ala-Ala-Ala
pH 7.5, 85°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.18
Ala-Ala
pH 7.5, 85°C
0.091 - 9.01
Ala-Ala-Ala
0.022
N-acetyl-L-alanyl 4-nitroanilide
pH 7.5, 85°C
0.116
N-acetyl-L-leucyl 4-nitroanilide
pH 7.5, 85°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8.3
pH 6.5: about 20% of maximal activity, pH 7.0: about 90% of maximal activity, pH 8.3: about 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
85 - 98
85°C: about 35% of maximal activity, 98°C: about 85% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
O58255_PYRHO
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
332
0
36924
TrEMBL
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36900
440000
gel filtration, HPLC
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 36900, wild-type enzyme, mutant enzymes D173N, E205Q and E206Q, SDS-PAGE
oligomer
x * 36900, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
sitting drop method, crystallized in the native and selenomethionine-substituted state, resolution of 3 A
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D173N
Km-value for Ala-Ala-Ala is not significantly different from the KM-value for the wild-type enzyme, kcat is significantly reduced
E205Q
Km-value for Ala-Ala-Ala is not significantly different from the KM-value for the wild-type enzyme, kcat is significantly reduced
E206Q
Km-value for Ala-Ala-Ala is not significantly different from the KM-value for the wild-type enzyme, kcat is significantly reduced
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
90
2 h, about 20% loss of activity
95
about 15% loss of activity after 1 h, about 70% loss of activity after 2 h
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
overexpressed in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Porciero, S.; Receveur-Brchot, V.; Mori, K.; Franzetti, B.; Roussel, A.
Expression, purification, crystallization and preliminary crystallographic analysis of a deblocking aminopeptidase from Pyrococcus horikoshii
Acta Crystallogr. Sect. F
61
239-242
2005
Pyrococcus horikoshii (O58255), Pyrococcus horikoshii (O59196), Pyrococcus horikoshii (O59485), Pyrococcus horikoshii
Manually annotated by BRENDA team
Onoe, S.; Ando, S.; Ataka, M.; Ishikawa, K.
Active site of deblocking aminopeptidase from Pyrococcus horikoshii
Biochem. Biophys. Res. Commun.
290
994-997
2002
Pyrococcus horikoshii (O58255), Pyrococcus horikoshii
Manually annotated by BRENDA team
Ando, S.; Ishikawa, K.; Ishida, H.; Kawarabayasi, Y.; Kikuchi, H.; Kosugi, Y.
Thermostable aminopeptidase from Pyrococcus horikoshii
FEBS Lett.
447
25-28
1999
Pyrococcus horikoshii (O58255), Pyrococcus horikoshii
Manually annotated by BRENDA team