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Information on EC 3.5.2.3 - dihydroorotase and Organism(s) Aquifex aeolicus and UniProt Accession O66990

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EC Tree
     3 Hydrolases
         3.5 Acting on carbon-nitrogen bonds, other than peptide bonds
             3.5.2 In cyclic amides
                3.5.2.3 dihydroorotase
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This record set is specific for:
Aquifex aeolicus
UNIPROT: O66990 not found.
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Word Map
The taxonomic range for the selected organisms is: Aquifex aeolicus
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
dihydroorotase, dho, dhoase, dihydroorotase domain, hudhoase, type i dhoase, human dhoase domain, type ii dho, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dihydroorotase
-
carbamoylaspartic dehydrase
-
-
-
-
DHOase
-
-
-
-
dihydroorotate dehydrolase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(S)-dihydroorotate + H2O = N-carbamoyl-L-aspartate
show the reaction diagram
complex formation of enzyme with aspartate transcarbamoylase drives reaction in the biosynthetic direction
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of cyclic amides
-
-
-
-
formation of cyclic amides
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -, -
SYSTEMATIC NAME
IUBMB Comments
(S)-dihydroorotate amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9024-93-5
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-dihydroorotate + H2O
N-carbamoyl-L-aspartate
show the reaction diagram
N-carbamoyl-L-aspartate
(S)-dihydroorotate + H2O
show the reaction diagram
-
-
-
r
N-carbamoyl-L-aspartate
dihydroorotate + H2O
show the reaction diagram
-
-
-
r
dihydroorotate + H2O
N-carbamoyl-L-aspartate
show the reaction diagram
-
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(S)-dihydroorotate + H2O
N-carbamoyl-L-aspartate
show the reaction diagram
N-carbamoyl-L-aspartate
(S)-dihydroorotate + H2O
show the reaction diagram
-
-
-
r
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
DHOase is a zinc-dependent metalloenzyme, analysis of the catalytic zinc environment, structure, overview. Cys181 is observed bonded to the zinc atom of the catalytic site, with residues 181-187 visible and tethered by the interaction of loop A with other regions of the protein. On one end, the 1200 bar zinc coordination is like in the monoclinic-ap form with the Cys181 recruited again by the zinc atom, instead of the water molecule; the Cys-181 sulfur atom rotates and becomes more and more closely connected to the zinc when pressure increases
Zn2+
-
one mol per mol of protein
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
DHCEDD
the peptide corresponds to the sequence 179DHCEDD185, and causes 50% inhibition of the wild-type enzyme
DHCEDDKLA
the peptide corresponds to the sequence 179DHCEDDKLA187, and causes 76% inhibition of the wild-type enzyme
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.24 - 24
N-carbamoyl-L-aspartate
3.03
dihydroorotate
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3 - 62.3
N-carbamoyl-L-aspartate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.28 - 38.3
N-carbamoyl-L-aspartate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 74
assay at
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
dihydroorotase (DHO) is an amidohydrolase that catalyzes the reversible condensation of carbamoyl aspartate to form dihydroorotate in de novo pyrimidine biosynthesis in virtually all organisms. Although the same reaction is catalyzed by all DHOs, the structure, oligomeric organization, and metal content of this family of enzymes is diverse
malfunction
the replacement of the zinc ligand Cys181 with glycine does not restore the latent catalytic activity suggesting that it plays a minor role in stabilizing loop A
metabolism
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
386000
-
gel filtration, enzyme-aspartate transcarbamoylase complex
49000
-
1 * 49000, SDS-PAGE
53000
-
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 43000, DHO, SDS-PAGE
hexamer
noncovalent association with aspartate transcarbamoylase
monomer
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
complex of aspartate transcarbamoylase and dihydroorotase
crystal structure analysis of the zinc environment at ambient pressure, at 600 bar, and at 1200 bar
hanging drop method, monoclinic structure has a novel cysteine ligand to the zinc that blocks the active site and functions as a cysteine switch, active site residues are located in disordered loops, which may function as a disorder-to-order entropy switch
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C181A
site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to wild-type
C181G
site-directed mutagenesis, the mutant does not restore the activity of DHOase that has been isolated from aspartate transcarbamylase (ATCase, EC 2.1.3.2)
D179V
site-directed mutagenesis, the mutant partially restores the activity of DHOase that has been isolated from aspartate transcarbamylase (ATCase, EC 2.1.3.2)
D183G
site-directed mutagenesis, the mutant partially restores the activity of DHOase that has been isolated from aspartate transcarbamylase (ATCase, EC 2.1.3.2)
E179V
site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to wild-type
E183V
site-directed mutagenesis, the mutant shows reduced activity and altered kinetics compared to wild-type
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
99
-
enzyme complex with aspartate transcarbamoylase, both activities are stable
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant isolated DHO and DDHO-ATC complex from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
His-tagged recombinant protein
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli strain BL21
gene pyrC, recombinant expression of wild-type and mutant DHO enzymes in Escherichia coli strain BL21(DE3), coexpression with pyrB encoding with aspartate transcarbamoylase (ATC, EC 2.1.3.2)
with a fusion peptide containing six histidine residues at the amino terminus
expression in Escherichia coli
-
expression in Escherichia coli, His-tag
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Purcarea, C.; Martin, P.; Vickrey, J.F.; Guy, H.I.; Edwards, B.F.; Evans, D.R.
Cloning, expression and preliminary X-ray analysis of the dihydroorotase from the hyperthermophilic eubacterium Aquifex aeolicus
Acta Crystallogr. Sect. D
58
154-156
2002
Aquifex aeolicus
Manually annotated by BRENDA team
Ahuja, A.; Purcarea, C.; Ebert, R.; Sadecki, S.; Guy, H.I.; Evans, D.R.
Aquifex aeolicus dihydroorotase: association with aspartate transcarbamoylase switches on catalytic activity
J. Biol. Chem.
279
53136-53144
2004
Aquifex aeolicus
Manually annotated by BRENDA team
Martin P.D; Purcarea, C.; Zhang, P.; Vaishnav, A.; Sadecki, S.; Guy-Evans, H.I.; Evans, D.R.; Edwards, B.F.
The crystal structure of a novel, latent dihydroorotase from Aquifex aeolicus at 1.7A resolution
J. Mol. Biol.
348
535-547
2005
Aquifex aeolicus (O66990), Aquifex aeolicus
Manually annotated by BRENDA team
Zhang, P.; Martin, P.D.; Purcarea, C.; Vaishnav, A.; Brunzelle, J.S.; Fernando, R.; Guy-Evans, H.I.; Evans, D.R.; Edwards, B.F.
Dihydroorotase from the hyperthermophile Aquifex aeolicus is activated by stoichiometric association with aspartate transcarbamoylase and forms a one-pot reactor for pyrimidine biosynthesis
Biochemistry
48
766-778
2009
Aquifex aeolicus (O66990), Aquifex aeolicus
Manually annotated by BRENDA team
Prange, T.; Girard, E.; Fourme, R.; Dhaussy, A.C.; Edwards, B.; Vaishnav, A.; Patel, C.; Guy-Evans, H.; Herve, G.; Evans, D.R.
Pressure-induced activation of latent dihydroorotase from Aquifex aeolicus as revealed by high pressure protein crystallography
FEBS J.
286
1204-1213
2019
Aquifex aeolicus (O66990), Aquifex aeolicus
Manually annotated by BRENDA team
Herve, G.; Evans, H.G.; Fernado, R.; Patel, C.; Hachem, F.; Evans, D.R.
Activation of latent dihydroorotase from Aquifex aeolicus by pressure
J. Biol. Chem.
292
629-637
2017
Aquifex aeolicus (O66990), Aquifex aeolicus
Manually annotated by BRENDA team