Information on EC 3.5.4.5 - cytidine deaminase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.5.4.5
-
RECOMMENDED NAME
GeneOntology No.
cytidine deaminase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2'-deoxycytidine + H2O = 2'-deoxyuridine + NH3
show the reaction diagram
(2)
-
-
-
cytidine + H2O = uridine + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amidine hydrolysis
Deamination
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Drug metabolism - other enzymes
-
-
Metabolic pathways
-
-
pyrimidine deoxyribonucleosides degradation
-
-
pyrimidine deoxyribonucleosides salvage
-
-
pyrimidine metabolism
-
-
Pyrimidine metabolism
-
-
pyrimidine ribonucleosides degradation
-
-
pyrimidine ribonucleosides salvage I
-
-
pyrimidine ribonucleosides salvage II
-
-
SYSTEMATIC NAME
IUBMB Comments
cytidine/2'-deoxycytidine aminohydrolase
Contains zinc. Catalyses the deamination of cytidine and 2'-deoxycytidine with similar efficiencies. The enzyme, which is widely distributed among organisms, is involved in salvage of both exogenous and endogenous cytidine and 2'-deoxycytidine for UMP synthesis.
CAS REGISTRY NUMBER
COMMENTARY hide
37259-56-6
-
9025-06-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Bacillus caldolyticus DSM405 (T53)
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
calf
-
-
Manually annotated by BRENDA team
variant Brassica oleracea capitata
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
guinea pig
-
-
Manually annotated by BRENDA team
pigeon
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
mosquito parasite
-
-
Manually annotated by BRENDA team
Escherichia coli 15 (ATCC 9723)
strain 15 (ATCC 9723)
-
-
Manually annotated by BRENDA team
strain A19
-
-
Manually annotated by BRENDA team
strain BL21(D3)
-
-
Manually annotated by BRENDA team
Escherichia coli DH5-alpha
strain DH5alpha
-
-
Manually annotated by BRENDA team
strain JF611
-
-
Manually annotated by BRENDA team
strain SO268
-
-
Manually annotated by BRENDA team
strain SO5201
-
-
Manually annotated by BRENDA team
strain Y-70-272
-
-
Manually annotated by BRENDA team
Felis catus domesticus
cat
-
-
Manually annotated by BRENDA team
chicken
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
rhesus monkey
-
-
Manually annotated by BRENDA team
no activity in Lactobacillus sp.
-
-
-
Manually annotated by BRENDA team
no activity in Moraxella sp.
no activity in Pseudomonas sp.
strain CT16
-
-
Manually annotated by BRENDA team
strain CT16
-
-
Manually annotated by BRENDA team
strain NRC-510, grown on nitrate
-
-
Manually annotated by BRENDA team
strain NRC-510, grown on nitrate
-
-
Manually annotated by BRENDA team
frog
-
-
Manually annotated by BRENDA team
rat
-
-
Manually annotated by BRENDA team
pig
-
-
Manually annotated by BRENDA team
human pathogen, Culbertson strain, epimastigotes, ATCC 30013
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
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
1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine + H2O
?
show the reaction diagram
1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)cytosine + H2O
?
show the reaction diagram
1-beta-D-arabinofuranosylcytosine + H2O
?
show the reaction diagram
2',2'-difluorodeoxycytidine + H2O
2',2'-difluorodeoxyuridine + NH3
show the reaction diagram
2',3'-dideoxycytidine + H2O
2',3'-dideoxyuridinel + NH3
show the reaction diagram
2',5'-anhydroarabinosylcytosine + H2O
2',5'-anhydroarabinosyluracil + NH3
show the reaction diagram
2'-deoxycytidine + H2O
2'-deoxyuridine + NH3
show the reaction diagram
2'-fluoro-2'-deoxy-5-iodocytosine arabinoside + H2O
?
show the reaction diagram
2'-fluoro-2'-deoxycytidine + H2O
2'-fluoro-2'-deoxyuridine + NH3
show the reaction diagram
2'-O-methylcytidine + H2O
?
show the reaction diagram
-
-
-
-
?
2'-thio-cytidine + H2O
2'-thiouridine + NH3
show the reaction diagram
-
-
-
-
?
3'-amino-2',3'-dideoxycytidine + H2O
3'-amino-2',3'-dideoxyuridine + NH3
show the reaction diagram
-
-
-
?
5'-bromoarabinosylcytosine + H2O
5'-bromoarabinosyluracil + NH3
show the reaction diagram
-
-
-
-
?
5'-chloroarabinosylcytosine + H2O
5'-chloroarabinosyluracil + NH3
show the reaction diagram
-
-
-
-
?
5'-chlorocyclocytidine + H2O
5'-chlorocyclouridine + NH3
show the reaction diagram
-
i.e. cytosine, 1beta-D-arabinofuranosyl-2,2'-anhydro-, hydrochloride
-
-
?
5'-deoxyfluorocytidine + H2O
5'-deoxy-5-fluorouridine + NH3
show the reaction diagram
-
-
-
-
?
5'-fluoro-5'-deoxycytidine + H2O
5'-fluoro-5'-deoxyuridine + NH3
show the reaction diagram
5'-methyl-2'-deoxycytidine + H2O
5'-methyl-2-deoxyuridine + NH3
show the reaction diagram
5,6-dihydrocytidine + H2O
5,6-dihydrouridine + NH3
show the reaction diagram
5-aza-2'-deoxycytidine + H2O
5-aza-2'-deoxyuridine + NH3
show the reaction diagram
5-azacytidine + H2O
5-azauridine + NH3
show the reaction diagram
5-azadeoxycytidine + H2O
5-azadeoxyuridine + NH3
show the reaction diagram
5-bromocytidine + H2O
5-bromouridine + NH3
show the reaction diagram
-
-
-
-
?
5-bromodeoxycytidine + H2O
5-bromodeoxyuridine + NH3
show the reaction diagram
-
-
-
-
?
5-chlorocytidine + H2O
? + NH3
show the reaction diagram
-
-
-
-
?
5-iodo-2'-deoxycytidine
5-iodo-2'-deoxyuridine + NH3
show the reaction diagram
-
-
-
-
?
5-iodocytidine + H2O
5-iodouridine + NH3
show the reaction diagram
5-iododeoxycytidine + H2O
5-iododeoxyuridine + NH3
show the reaction diagram
5-methyl-dCMP + H2O
5-methyl-dCyd
show the reaction diagram
5-methylcytidine + H2O
5-methyluridine + NH3
show the reaction diagram
6-azacytidine + H2O
6-azauridine + NH3
show the reaction diagram
-
-
-
-
?
bromodeoxycytidine + H2O
bromodeoxyuridine + NH3
show the reaction diagram
-
-
-
?
CTP + H2O
UTP + NH3
show the reaction diagram
cytidine + H2O
uridine + NH3
show the reaction diagram
cytosine arabinoside + H2O
?
show the reaction diagram
cytosine arabinoside + H2O
uridine arabinoside + NH3
show the reaction diagram
cytosine beta-D-arabinofuranoside + H2O
?
show the reaction diagram
-
-
-
-
?
dCTP + H2O
dUTP + NH3
show the reaction diagram
deoxycytidine + H2O
deoxyuridine + NH3
show the reaction diagram
gemcitabine + H2O
2'-deoxy-2',2'-difluorouridine
show the reaction diagram
-
CDA is the major enzyme involved in gemcitabine inactivation, a cytotoxic drug commonly used in the treatment of pancreas and non-small cell lung cancer
-
-
-
N-4-monoacetylarabinosylcytosine
uracil arabinoside + carbamate
show the reaction diagram
N4-methylcytidine + H2O
uridine + methylamine
show the reaction diagram
-
-
-
-
r
additional information
?
-
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
2'-deoxycytidine + H2O
2'-deoxyuridine + NH3
show the reaction diagram
5-aza-2'-deoxycytidine + H2O
5-aza-2'-deoxyuridine + NH3
show the reaction diagram
-
5-AZA-CdR shows an antineoplastic action on 3T3 cells and V5 cells transduced with CR deaminase gene, pharmakokinetic analysis in mice, overview
-
-
?
cytidine + H2O
uridine + NH3
show the reaction diagram
cytosine arabinoside + H2O
?
show the reaction diagram
-
-
-
?
cytosine beta-D-arabinofuranoside + H2O
?
show the reaction diagram
-
-
-
-
?
deoxycytidine + H2O
deoxyuridine + NH3
show the reaction diagram
additional information
?
-
-
cytidine deaminase plays an important role in the activation of the anticancer drug capesitabine in the human body
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zinc
-
wild-type enzyme and mutant enzymes R56A, R56Q and C53H/R56Q contain about 1 mol of zinc per subunit. Mutant enzyme R56D contains 0.2 mol of zinc per subunit
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(cytidyl)acryloylaminohexanoic acid
-
i.e. CV6
1,10-phenanthroline
-
-
1-(beta-D-ribofuranosyl)-2-pyrimidone
1-(beta-D-ribofuranosyl)-dihydropyrimidine-2-one
1-methyladenosine
-
competitive inhibition
2'-deoxyuridine
-
-
2'-deoxyzebularine 5'-monophosphate
-
-
2-beta-D-ribofuranosyl-1,2,4-triazol-3-one
-
-
2-mercaptoethanol
-
-
2-thio-6-azarudine
-
-
3'-azido-3'-deoxythymidine
-
-
3,4,5,6,tetrahydrozebularine
3,4,5,6-Tetrahydrouridine
3,4-dihydrouridine
3,4-dihydrozebularine
3-deazauridine
4-amino-1-(5-hydroxypentyl)pyrimidin-2(1H)-one
-
-
4-amino-1-hexyl-5,6-dihydropyrimidin-2(1H)-one
-
-
4-hydroxy-1-(beta-D-ribofuranosyl)piperidin-2-one
-
-
5,6-Dihydrouridine
5-(Chloromercuri)cytidine
5-bromodeoxyuridine
5-Bromouridine
-
-
5-fluoro-2'-deoxyuridine
5-fluoropyrimidin-2-one ribonucleoside
-
-
5-fluorouridine
5-fluorozebularine
5-iodo-2'-deoxyuridine
-
-
5-methyluridine
-
-
5-{(1E)-3-[(5-carboxypentyl)amino]-3-oxoprop-1-en-1-yl}-5,6-dihydrocytidine
6-azauridine
-
-
6-Thioguanine
-
-
6-[3-(5-cytidyl)acryloylamino]hexanoic acid
-
i.e. CV6
acetone
-
-
adenosine
ara-U
-
-
blasticidin S
-
-
CuSO4
-
complete inhibition
cytidine
-
strong substrate inhibition above 0.5 mM
cytidine monophosphate
-
-
deoxyadenosine
deoxyguanosine
deoxyuridine
diazepinone riboside
dihydrothymidine
-
competitive inhibition
dipicolinic acid
-
-
dithiothreitol
-
-
dTMP
-
0.400 mM 50% inhibition
GTP
-
0.750 mM 85% inhibition
guanosine
HgCl2
-
complete inhibition
mercaptoethanol
-
-
Mersalyl acid
N-ethylmaleimide
p-chloromercuribenzoate
p-mercuribenzoate
phosphapyrimidine
-
-
phosphapyrimidine nucleoside
-
-
pseudouridine
-
-
SDS
-
wild-type tetramer dissociates into enzymatically inactive monomers, without intermediate forms via a non-cooperative transition. Extensive dialysis or dilution of the inativated monomers restores completely the activity. 5-Fluorozebularine disfavours dissociation of the tetramer into subunits in the wild-type enzyme, but not in the mutant enzyme F137W/W113F
tetrahydrouridine
thioglycolic acid
-
0.7 mM 88% relative activity, complete inhibition at 7 mM
thymidine
thymine riboside
uracil arabinoside
-
4.0 mM, 48% inhibition
Urea
-
above 2 M, reversible inactivation
uridine
zebularine
-
inhibits the enzyme, but also DNA methylation, competitive versus 5-aza-2'-deoxycytidine, zebularine affects the antineoplastic action of 5-AZA-CdR on 3T3 cells and V5 cells transduced with CR deaminase gene, overview
Zn2+
-
above 10 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5-Bromouridine
-
-
bromodeoxyuridine
-
-
cytidine
dithiothreitol
-
-
iodoacetic acid
-
0.10 mM, activity 105%
N-ethylmaleimide
-
0.10-0.20 mM, activity 114%
p-chloromercuribenzoate
-
0.105 mM, activity 112%
uridine
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.31 - 3.9
1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine
0.33 - 0.66
1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)cytosine
0.27 - 0.51
1-beta-D-arabinofuranosylcytosine
0.0092 - 0.0957
2',2'-difluorodeoxycytidine
0.0726
2',2'-difluorodeoxyuridine
-
i.e. dFdU, variation of plasma ranges between 24.9 and 72.6 microM
0.023 - 1.059
2'-deoxycytidine
0.07
2'-fluoro-2'-deoxy-5-iodocytosine arabinoside
-
-
0.091
2-deoxycytidine
-
-
0.087
5'-fluoro-5'-deoxycytidine
-
-
0.11 - 0.113
5,6-Dihydrocytidine
0.0477
5-aza-2'-deoxycytidine
-
pH 7.4, 37C
0.058 - 2.27
5-azacytidine
0.087 - 0.1
5-iododeoxycytidine
0.13
5-methyl-dCMP
-
pH 7.0, 37C
0.04 - 1.25
5-methylcytidine
0.06
5-Methyldeoxycytidine
-
-
4.2
6-azacytidine
-
-
0.14
CMP
-
pH 7.0, 37C
0.0092 - 6.6
cytidine
0.089
cytosine
-
-
0.058 - 0.385
cytosine arabinoside
1.475
cytosine beta-D-arabinofuranoside
-
at pH 7.5 and 27C
-
0.089
cytosine deoxyriboside
-
-
0.055
dCMP
-
pH 7.0, 37C
0.0075 - 8.53
deoxycytidine
additional information
additional information
-
kinetics of wild-type and mutant enzymes
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.5
2'-deoxycytidine
Mycobacterium tuberculosis
-
in 50 mM Tris-HCl buffer pH 7.5 at 25C
4.8
cytidine
Mycobacterium tuberculosis
-
in 50 mM Tris-HCl buffer pH 7.5 at 25C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.034 - 0.045
(cytidyl)acryloylaminohexanoic acid
0.00042 - 0.008
3,4,5,6-Tetrahydrouridine
0.015 - 0.04
5,6-Dihydrouridine
0.00006 - 0.0017
5-fluorozebularine
0.015 - 0.045
6-[3-(5-cytidyl)acryloylamino]hexanoic acid
0.3
CMP
0.39
cytidine monophosphate
-
tetramer of recombinant subunit CDA1, 37C
0.000025
diazepinone riboside
-
-
0.1 - 0.4
uridine
0.00095
zebularine
-
pH 7.4, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00164
-
-
0.0059
-
-
0.011
-
-
0.536
-
activity in non-small cell lung cancer patients with CDA Lys27Lys protein, in 50 mM beta-mercaptoethanol, 0.1 M Tris-HCl, pH 8.0, substrate 0.25 mM gemcitabine
0.7254
-
activity in non-small cell lung cancer patients carrying the Lys27Gln polymorphism and the Gln27Gln polymorphism, in 50 mM beta-mercaptoethanol, 0.1 M Tris-HCl, pH 8.0, substrate 0.25 mM gemcitabine
1.68
-
crude extract, pH 7.5, at 25C, using 2'-deoxycytidine as substrate
2.35
-
crude extract, pH 7.5, at 25C, using cytidine as substrate
3.75
-
after 2.2fold purification, pH 7.5, at 25C, using 2'-deoxycytidine as substrate
3.96
-
sarcoma tumour
4
-
amplified gene, 5'-aza-2'-deoxycytidine as substrate
4.9
-
amplified gene, 2'-thiocytidine as substrate
5.44
-
after 2.3fold purification, pH 7.5, at 25C, using cytidine as substrate
23.9
-
amplified gene, 5'-methylcytidine as substrate
29
-
colon tumour
31.8
-
amplified gene, ara-cytidine as substrate
43
-
amplified gene, cytidine as substrate
47.9
-
amplified gene, 5'-methyl-2'-deoxycytidine as substrate
64.1
-
-
68.4
-
amplified gene, 2'-fluoro-2'-deoxycytidine as substrate
86.7
-
pancreas tumour
97.6
-
amplified gene, deoxycytidine as substrate
225
-
pH 7.0, 37C
738
-
ovarian tumour
3558
-
amplified gene
additional information
-
variation between 1.51 to 5.50 nmol/h/mg protein, about 100-fold lower than that in liver
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
-
-
5.9 - 7.35
-
deamination rate 50% higher in phosphate buffers with pH 7.25-8.7
6 - 7.5
-
-
6.5 - 8.5
-
-
7.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 10.5
-
deoxycytidine as substrate, specific activity constant in this range, beyond these values irreversible inactivation occurs
3.5 - 10.5
-
deoxycytidine as substrate, beyond these values irreversible inactivation occurs
3.8 - 7.5
6 - 11
6.5 - 10.7
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 62
-
mutant Y60G shows an unusual behavior with a mean of 75% activity for temperatures within this range and three peaks of maximum activity at about 30C, 46C, and 62C
58
-
recombinant wild-type enzyme
99
-
maximum of activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 70
6C: 45% of maximal activity, 70C: 35% of maximal activity
12 - 66
-
12C: about 55% of maximal activity, 66C: about 65% of maximal activity
15 - 65
-
mutant Y60G shows an unusual behavior with a mean of 75% activity for temperatures within this range and three peaks of maximum activity at about 30C, 46C, and 62C
34 - 74
34C: 45% of maximal activity, 74C: 70% of maximal activity
37 - 75
-
60% of maximal activity at 75C for the wild-type enzyme, 35% for mutant R68Q
40 - 62
-
heat treatment for 5 min at 50-60C completely abolishes CMP and substrate inhibition
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
tetramer of recombinant subunit CDA2, isoelectric focusing
4.55 - 4.9
-
5 different isoforms of native enzyme, isoelectric focusing
4.9
chromatofocusing
5
-
tetramer of recombinant subunit CDA1, isoelectric focusing
7.3
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
glioblastoma multiforme (GBM) patients
Manually annotated by BRENDA team
-
glioblastoma multiforme (GBM) patients
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
CDA is partially concentrated in the nuclei
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Burkholderia pseudomallei (strain 1710b)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Mycobacterium marinum (strain ATCC BAA-535 / M)
Mycobacterium smegmatis (strain ATCC 700084 / mc(2)155)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
16170
-
deduced from cDNA nucleotide sequence
32500
-
calculated from cDNA open reading frame
32560
-
calculated from cDNA open reading frame
32580
-
calculated from amino acid sequence
45000
-
gel filtration
51000
-
human granulocytes
52990
-
gel filtration
60000
-
gel filtration
62000
-
gel filtration, native enzyme
63000
-
gel filtration
66000
-
gel filtration, SDS-PAGE
73000
-
gel filtration, nonhomogenous preparation
77000 - 78000
-
gel filtration
90000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 20800, SDS-PAGE
homotetramer
monomer
tetramer
additional information
-
wild-type tetramer dissociates into enzymatically inactive monomers, without intermediate forms via a non-cooperative transition. Extensive dialysis or dilution of the inativated mponomers restores completely the activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of tetrameric cytidine deaminase at 2.0 A resolution, hanging drop vapour-diffusion method
-
hanging drop vapour diffusion method. R56A and R56Q crystallize in the same space group as the wild-type enzyme with two subunits in the asymmetric unit, whereas C53H/R56Q can not crystallize in this crystal form but is crystallized in another space group, P3(2)21, with a full tetramer in the asymmetric unit
-
comparison of theoretically predicited model and experimentally elucidated structure
-
in complex with inhibitor diazepinone riboside. Inhibitor is able to establish a canonical pi/pi interaction with key active site residue F137
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in complex with tetrahydrouridine, 3-deazauridine, or cytidine. Two alternate conformations of R68 influence zinc-product interaction; purified recombinant CDA complexed with either tetrahydrouridine, 3-deazauridine, or cytidine, hanging drop vapor diffusion technique, 25C, protein solution, containing 10-15 mg/ml protein in 20 mM Tris-HCl, pH 7.5, 1 mM DTT, and 5 mM ligand, is mixed with crystallization solution containing 2.0 M ammonium sulfate and 0.1 M Tris-HCl, pH 8.5, crystals appear after a few hours or several days, respectively, X-ray diffraction structure determination and analysis at 1.48-2.36 A resolutions, molecular replacement
hanging drop vapor diffusion method
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purified CDA in complex with uridine and deoxyuridine, hanging drop vapor diffusion method, 0.002 ml of 12 mg/ml protein in 20 mM Tris-HCl pH 7.5 is mixed with 0.002 ml of reservoir solution containing 0.1 M HEPES, pH 7.5 and 4.3 M sodium chloride, ligands uridine and deoxyuridine are added by soaking method, X-ray diffractiuon structure determination and analysis at 2.4 and 1.9 A resolution,molecular dynamics simulation, structure modeling
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pH STABILITY
ORGANISM