Information on EC 3.5.4.12 - dCMP deaminase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.5.4.12
-
RECOMMENDED NAME
GeneOntology No.
dCMP deaminase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
dCMP + H2O = dUMP + NH3
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of amidines
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
pyrimidine deoxyribonucleotides biosynthesis from CTP
-
-
pyrimidine metabolism
-
-
Pyrimidine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
dCMP aminohydrolase
Also acts on some 5-substituted dCMPs.
CAS REGISTRY NUMBER
COMMENTARY hide
9026-92-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain ED40
-
-
Manually annotated by BRENDA team
host Bacillus subtilis
-
-
Manually annotated by BRENDA team
bacteriophage S-TIM5
-
UniProt
Manually annotated by BRENDA team
bacteriophage SP8
host Bacillus subtilis
-
-
Manually annotated by BRENDA team
T2-, T4-, and T6-phage induced enzyme in Escherichia coli
-
-
Manually annotated by BRENDA team
Herpes simplex virus
Ehrlich's ascites tumor cells and various gastrointestinal tumors
-
-
Manually annotated by BRENDA team
subsp. mycoides
-
-
Manually annotated by BRENDA team
no activity in Escherichia coli
no activity in Salmonella typhimurium
cultivar Zhonghua11
-
-
Manually annotated by BRENDA team
i.e. PBCV-1 , a chlorovirus from host Chlorella sp. strain NC64A
SwissProt
Manually annotated by BRENDA team
induced by
-
-
Manually annotated by BRENDA team
Sea urchin
-
-
-
Manually annotated by BRENDA team
induced by
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
dCMPD is pivotal in deoxyuridine analogs nucleotide formation
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(+)-FTCMP + H2O
?
show the reaction diagram
-
-
-
-
?
2',2'-difluoro-dCMP + H2O
2',2'-difluoro-dUMP + NH3
show the reaction diagram
-
-
-
-
?
2',3'-dideoxy-5-fluoro-3'-thiacytidine + H2O
?
show the reaction diagram
-
good substrate
-
-
?
5-aza-dCMP + H2O
5-aza-dUMP + NH3
show the reaction diagram
-
-
-
-
?
5-bromo-dCMP + H2O
5-bromo-dUMP + NH3
show the reaction diagram
5-chloro-dCMP + H2O
5-chloro-dUMP + NH3
show the reaction diagram
5-fluoro-dCMP + H2O
5-fluoro-dUMP + NH3
show the reaction diagram
5-Hg-dCMP + H2O
5-Hg-dUMP + NH3
show the reaction diagram
5-hydroxymethyl-dCMP + H2O
5-hydroxymethyl-dUMP + NH3
show the reaction diagram
5-iodo-dCMP + H2O
5-iodo-dUMP + NH3
show the reaction diagram
5-methyl-dCMP + H2O
5-methyl-dUMP + NH3
show the reaction diagram
5-methyl-dCMP + H2O
dTMP + NH3
show the reaction diagram
ara-CMP + H2O
ara-UMP + NH3
show the reaction diagram
beta-D-2',2'-difluorodeoxycytidine + H2O
beta-D-2',2'-difluorodeoxyuridine + NH3
show the reaction diagram
-
-
-
-
?
CMP + H2O
UMP + NH3
show the reaction diagram
dCMP + H2O
dUMP + NH3
show the reaction diagram
dCMP-Hg-S-CH2-CH2-OH + H2O
dUMP-Hg-S-CH2-CH2-OH + NH3
show the reaction diagram
-
better substrate of T form: in the presence of dTTP
-
-
?
dCTP + H2O
dUTP + NH3
show the reaction diagram
-
-
-
?
deoxycytidine + H2O
deoxyuridine + NH3
show the reaction diagram
PSI-6130-MP + H2O
?
show the reaction diagram
-
weak substrate
-
-
?
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
5-methyl-dCMP + H2O
dTMP + NH3
show the reaction diagram
-
deamination pathway, overview
-
-
?
dCMP + H2O
dUMP + NH3
show the reaction diagram
dCTP + H2O
dUTP + NH3
show the reaction diagram
O41078
-
-
-
?
deoxycytidine + H2O
deoxyuridine + NH3
show the reaction diagram
-
the enzyme is a key enzyme in the inactivation of deoxycytidine and several chemotherapeutically important nucleoside analogues
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
no coenzyme required
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Divalent cations
metal ion
-
required, in the absence of metal ions: 40% activity, Zn2+ highest activity, around 20% activity with Co2+, Mn2+, and Ni2+, no activity in the presence of Mg2+, Ca2+ and Fe2+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(4Z,7Z,10Z,13Z,16Z,19Z)-henicosa-4,7,10,13,16,19-hexaenoic acid
-
decreases the enzyme activity in vivo in transformed NM-16 fibroblasts, but increases it in normal Rat-2 fibroblasts
2',2'-difluoro-dCTP
-
dual mechanism of inhibition; partial purified enzyme
2',2'-difluorodeoxycytidine
-
high cellular nucleotide levels: inhibition: Concentration-dependent inhibition; inhibition by nucleotides of 2',2'-difluorodeoxycytidine, dual mechanism of inhibition
2',3'-di-dTTP
-
7% inhibition
2',3'-dihydro-2',3'-dideoxy-thymidine monophosphate
-
-
2'-beta-D-deoxyribose-pyrimidin-2-one 5'-phosphate
-
competitive inhibition, Ki: 0.000012 mM
2'-deoxythymidine
-
0.05 mM: 60% inhibition, 0.01 mM: 5% inhibition; dThd
3'-azido-2',3'-dideoxy-thymidine monophosphate
-
-
3,4,5,6-tetrahydro-2'-deoxyuridine
3,4,5,6-tetrahydro-2'-dUMP
5-(acryloylamino-pentanol-1)2'-beta-D-dUMP
-
-
-
5-bromo-dUTP
-
92% inhibition
5-fluoro-dUMP
-
-
5-Hg-dCMP
5-Hg-dUMP
-
potent inhibitor, 0.0001 mM: 54% inhibition
ADP
-
slight inhibition
AMP
-
slight inhibition, competitive inhibitor, mechanism of inhibition
arabinosylcytosine
-
ara-C, deactivating dCMP deaminase
beta-D-2',2'-difluorodeoxycytidine
-
competitive
beta-L-2',3'-dideoxy-3'-thiacytidine monophosphate
-
slight inhibition
beta-L-2',3'-dideoxy-5'-fluoro-3'-thiacytidine monophosphate
-
-
Co2+
-
1 mM: 75% inhibition
Cu2+
-
1 mM: 98% inhibition
D-beta-2'-fluoro-5-methyl-arabinofuranosyluracil monophosphate
-
no inhibition by L-beta-2'-fluoro-5-methyl-arabinofuranosyluracil monophosphate
deoxytetrahydrouridine
-
-
deoxythymidine 5'-triphosphate
-
allosteric feedback regulation of the enzyme by products of the metabolic pathway, allosteric regulation involved residues 112 and 115
dGDP
-
slight inhibition
Glutaraldehyde
glycerol
-
substrate dCMP: activation, substrate dCMP-Hg-S-CH2-CH2-OH, 20% glycerol: inhibition, removed by dTTP
guanidine hydrochloride
-
0.5 M: 50% inhibition, 1 M: 100% inhibition
H4dUMP
inhibits both dCTP and dCMP deaminase activities
Herpes antiserum
Herpes simplex virus
-
inhibition of Herpes infected-cell enzyme
-
Hg(CH3COO)2
-
Hg(acetate)2; mercuroacetate, potent inhibitor, 0.0002 mM: 26% inhibition, 0.001 mM: 40% inhibition
PDRP
inhibits both dCTP and dCMP deaminase activities
pre-immune serum
Herpes simplex virus
-
inhibition of Herpes infected-cell enzyme
-
pyrimidin-2-one deoxyribotide
-
active site binding structure, overview
TDP
-
slight inhibition, competitive inhibitor, mechanism of inhibition
tetrahydrodeoxyuridine
-
-
UTP
-
slight inhibition
UTP-adipic hydrazide
-
cooperative inhibition, reversed by dCTP
zebularine
-
-
Zn2+
-
1 mM: 94% inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(4Z,7Z,10Z,13Z,16Z,19Z)-henicosa-4,7,10,13,16,19-hexaenoic acid
-
increases the enzyme activity in vivo in normal Rat-2 fibroblasts, but decreases it in transformed NM-16 fibroblasts
1-beta-D-arabinofuranosylcytosine 5'-triphosphate
-
low activation, allosteric activator
2-mercaptoethanol
-
enzyme requires dCTP, Zn2+, and 2-mercaptoethanol
3,4,5,6-tetrahydro-2'-dUMP
-
high-affinity inhibitor, rapid, reversible inhibition, kinetics of inhibition. At low substrate or competitor concentrations: activation. Tetrahydro-dUMP activates enzyme at low substrate concentrations
5-hydroxymethyl-dCTP
ammonium sulfate
-
substrate dCMP-Hg-S-CH2-CH2-OH, 0.2 M ammonium sulfate: activation, reversed by cCTP
CDP
-
low activation, allosteric activator
dCMP
-
allosterically increases dCMPD activity
deoxycytidine 5'-triphosphate
-
allosteric feedback regulation of the enzyme by products of the metabolic pathway, allosteric regulation involved residues 112 and 115
dGTP
-
very slight activation
dUTP
-
89% activation
Glutaraldehyde
-
substrate dCMP: rapid inhibition, protection by dGMP, or dTTP or both is slight, substrate dCMP-Hg-S-CH2-CH2-OH: 3fold activation
glycerol
-
substrate dCMP: activation, substrate dCMP-Hg-S-CH2-CH2-OH, 20% glycerol: inhibition
Herpes antiserum
-
activation of host-cell enzyme
-
pre-immune serum
-
activation of host-cell enzyme, twice as effective as Herpes antiserum
-
TTP
-
activating enzyme up to 1.7fold at concentration of 0.00025 mM, but inhibiting at higher concentration, 0.015 mM: 85% inhibition
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
5-methyl-dCMP
-
pH 8.3, in the presence of dCTP or 5-hydroxymethyl-dCTP. Decreasing the pH to 6.0 lowers the apparent Km
0.5 - 12
CMP
0.021 - 1
dCMP
0.26
dCMP-Hg-S-CH2-CH2-OH
-
-
-
0.347 - 0.467
deoxycytidine
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
24.9 - 1400
dCMP
5.7
dCTP
Paramecium bursaria Chlorella virus 1
O41078
pH 9.5, 42C
additional information
additional information
Enterobacteria phage T4
-
mutants R115E and R115Q possessing turnover number or kcat that is about 15% that of wild-type enzyme
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0394
2',3'-dihydro-2',3'-dideoxy-thymidine monophosphate
-
pH 7.5
0.0367
3'-azido-2',3'-dideoxy-thymidine monophosphate
-
pH 7.5
0.0142
5-fluoro-dUMP
-
pH 7.5
0.0103
D-beta-2'-fluoro-5-methyl-arabinofuranosyluracil monophosphate
-
pH 7.5
0.0257
dUMP
-
pH 7.5
0.0046
TMP
-
pH 7.5
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.37
-
-
23
-
substrate: dCMP-Hg-S-CH2-CH2-OH
90 - 100
-
mutant R115E
121
-
mutant R115Q
500
-
HeLa cells
820
-
substrate: dCMP
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3
-
in the absence of dCTP
6.5 - 7.3
-
broad pH optimum
7.2
-
assay at
7.5
-
assay at
7.5 - 8.5
-
HeLa cells, in presence or absence of dCTP and Mg2+
7.5
-
assay at
7.5 - 9.5
-
broad pH optimum
8.1
-
assay at
8.3
-
in the presence of dCTP or 5-hydroxymethyl-dCTP
8.4
-
BHK-21/C13 cells
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
bacteriophage S-TIM5
more than 80% activity between pH 6.0 and 8.0
7 - 8.5
-
50% maximal activity at pH 7.0 and 8.5
7 - 9
-
pH 7.0: 60%, pH 8.2: 98% and pH 9.0 96% of maximum activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38
-
assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
baby-hamster kidney
Manually annotated by BRENDA team
Sea urchin
-
-
Manually annotated by BRENDA team
-
the enzyme activity is higher in normal Rat-2 fibroblast compared to transformed NM-16 cells
Manually annotated by BRENDA team
-
quiescent cells infected with human cytomegalovirus, HCMV
Manually annotated by BRENDA team
-
BHK-21/C13 cells, baby-hamster kidney cells
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
transformed fibroblast cell line
Manually annotated by BRENDA team
-
fibroblast cell line
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
Streptococcus mutans serotype c (strain ATCC 700610 / UA159)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38200 - 113000
-
recombinant mutant F112A, gel filtration, dependent on protein concentration, in presence and absence of fdCTP
39800 - 118000
-
recombinant mutant R115Q, gel filtration, dependent on protein concentration, in presence and absence of fdCTP
41600 - 45400
-
recombinant mutant R115E, gel filtration, dependent on protein concentration, in presence and absence of fdCTP
44300
-
R115E, HPLC gel filtration
109500
-
gel filtration
110000 - 122000
-
recombinant wild-type enzyme, gel filtration, dependent on protein concentration, in presence and absence of fdCTP
111900
-
F112A, HPLC gel filtration
113100
-
amino acid analysis
115000
-
BHK-21/C13 cells, sucrose density gradient centrifugation
117000
-
in the presence of dCTP and Mg2+, 4C, gel filtration
120000
124000
-
PAGE and sedimentation equilibrium
124500
-
R115Q, HPLC gel filtration
127000
-
wild-type, HPLC gel filtration
130000
-
BHK-21/C13 cells, gel filtration
170000
-
gel filtration and sucrose density gradient centrifugation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
bound to dTTP, hanging drop vapor diffusion method, using 0.2 M potassium formate, 20% (w/v) PEG 3350
bacteriophage S-TIM5
purified mutant R115E free or in complex with active site binding inhibitor pyrimidin-2-one deoxyribotide, the mutant R115E is dimeric in solution, while hexameric in crystals, hanging drop vapour diffusion method, 0.28 mM mutant R115E protein in 20 mM phosphate, pH 7.2, 0.38 mM inhibitor, and 30 mM DTT, is mixed with an equal volume of reservoir solution containing 1.3 M ammonium sulfate, 0.1 M sodium citrate, pH 6.0, and 10 mM DTT, 22C, 3 weeks, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
dCMP deaminase in complex with dCTP and an intermediate analogue, X-ray diffraction structure determination and analysis at 1.66-3.0 A resolution, the crystal structure of the free-state enzyme is determined by the zinc multiwavelength anomalous dispersion, MAD, method
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0
-
half-life: 30 min, Zn2+, thiols, 2-mercaptoethanol, and dCTP protect against inactivation
43
-
in the presence of ethyleneglycol: 100% stable
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol stabilizes
dCTP and Mg2+ stabilize
-
dCTP and MgCl2 stabilize
-
dCTP or glycerol stabilize
dCTP stabilizes
dCTP stabilizing hexameric state, dTTP destabilizing
dCTP, 0.125 mM, or ethyleneglycol, 20% v/v, stabilize
-
dCTP, or to a lesser extent dTTP, stabilize against heat inactivation, protein denaturants and proteolytic enzymes
-
dithiothreitol, 2 mM, stabilizes
-
EDTA: denaturation, EDTA removes the two resident Zn2+ atoms /subunit
-
enzyme extremely labile, stabilized by dCTP /Mg2+
-
enzyme is unstable
enzyme very heat-labile, stabilized against heat inactivation by dCTP, 0.125 mM and ethyleneglycol, 20% v/v
-
enzyme very sensitive to preliminary heating, dCTP, 0.125 mM, and Mg2+, 2 mM, stabilize against heat inactivation
ethyleneglycol prevents protein denaturation during freezing in the presence of 2-mercaptoethanol
-
ethyleneglycol, 10-20% stabilizes
-
ethyleneglycol, 20%, stabilizes against thermal and UV inactivation and denaturation factors with different mechanism of action
-
glycerol stabilizes
glycerol, 15%, stabilizes
-
guanidine hydrochloride, 1 M, complete loss in activity accompanied by disaggregation of enzyme, 6 M: denaturation, complete reactivation with 50 mM 2-mercaptoethanol
-
guanidine-HCl, 6 M, denaturation, complete restoration of activity on removal of denaturant by dilution or dialysis
-
highly stable enzyme
-
some free nucleotides, most effective dCTP, stabilize
-
thiols stabilize
thiols, such as 2-mercaptoethanol or dithiothreitol, stabilize, in the absence of thiols the enzyme is highly unstable
-
Zn2+ or dCTP stabilize, protect against inactivation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.1 M Na-phosphate, pH 7.5, 50% glycerol, 12 months, no activity lost
-
-20C, 10-20% ethyleneglycol, 0.01 mM dCTP, 1 mM MgCl2, 20 mM 2-mercaptoethanol and 0.05% Triton X-100, 1 year, no loss in activity
-
-40C, solid ammonium sulfate to 80% saturation, stable for longer periods of storage
-
-50C, 0.05 mM dCTP, 1 mM MgCl2 or 2 mM dithiothreitol, 18 h, 40% activity lost
-
-70C, 20% v/v ethyleneglycol, stable for 2 months
-
-70C, pH 7.8, 2 mM 2-mercaptoethanol
-
0-4C, 0.1 M 2-mercaptoethanol, stable for extended periods
-
0-4C, neutral pH, 0.1 M 2-mercaptoethanol, stable for several months
-
0C, 2-mercaptoethanol, stable for more than 6 h
-
0C, half-life: 30 min
-
frozen in absence of 2-mercaptoethanol, indefinitely stable
-
on ice, 0.05 M Tris, pH 8.0, 20 mM dithiothreitol, stable for weeks
-
the purified recombinant His-tagged enzyme in stable without precipitation for more than 1 year in 50% glycerol
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
BHK-21/C13 cells , enzyme from non-infected cells and cells infected by Herpes simplex
-
cloned wild-type and mutant F112A
-
HeLa cells
-
HisPur cobalt resin column chromatography and Superdex 200 gel filtration
bacteriophage S-TIM5
host baby-hamster, BHK-21/C13 cells
Herpes simplex virus
-
native enzyme 60fold from Hep-G2 cells by adsorption chromatography
-
native enzyme from sperm cells by ion exchange chromatography and preparative SDS-PAGE
-
partial
partial, BHK-21/C13 cells, baby-hamster
-
partial, Ehrlich's ascites tumor cells
-
partial, leukemia cells
-
partial, strain ACM-13
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
recombinant His6-tagged enzyme from Escherichia coli by nickel affinity chromatography, the protein quickly precipitates at concentration higher than 2 mg/ml
recombinant mutant R115E from Escherichia coli
-
recombinant wild-type and mutant enzymes from Escherichia coli; wild-type and mutants R115E, R115Q and F112A
-
T2-phage infected Escherichia coli
-
T4-phage infected Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA from HeLa cells can be expressed in an active form
-
cloned and amplified in a high-expression system, expression in Escherichia coli strain BL21-DE3/pLysS
-
cloned and overexpressed in Escherichia coli strain BL21-DE3/pLysS
-
expressed in Escherichia coli BL21(DE3) cells
bacteriophage S-TIM5
expressed in onion epidermal cells and rice protoplasts
-
expression of mutant R115E in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli
-
ORF A596R, DNA and amino acid sequence determination and analysis, expression of the His6-tagged enzyme in Escherichia coli strain DH5MCR, phylogenetic analysis
overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
N43G
bacteriophage S-TIM5
the mutation results in a dramatic loss of activity with less than 20% activity compared to the wild type
T61A
bacteriophage S-TIM5
the mutant shows about 65% activity compared to the wild type enzyme
T61D
bacteriophage S-TIM5
the mutant shows about 7% activity compared to the wild type enzyme
T61S
bacteriophage S-TIM5
the mutant shows about 145% activity compared to the wild type enzyme
T61V
bacteriophage S-TIM5
the mutant shows about 50% activity compared to the wild type enzyme
W42A
bacteriophage S-TIM5
inactive
W42F
bacteriophage S-TIM5
the mutant shows about 118% activity compared to the wild type enzyme
W42Y
bacteriophage S-TIM5
the mutant shows about wild type activity
Y116E
bacteriophage S-TIM5
inactive
Y116F
bacteriophage S-TIM5
the mutant shows about 140% activity compared to the wild type enzyme
R115Q
-
mutant enzyme active in the absence of dCTP and Mg2+, possessing turnover number or kcat that is about 15% that of wild-type enzyme, specific activity about 40-50% of wild-type enzyme. Molecular weight analysis using HPLC size gel filtration in the presence of SDS and dCTP: wild-type is a hexamer, R115Q varies from hexamer to dimer; site-directed mutagenesis, the mutant enzyme shows reduced turnover and activity compared to the wild-type enzyme, but no longer requires deoxycytidine 5-triphosphate for activation in contrast to the wild-type enzyme
S178F
-
site-directed mutagenesis, the mutation is responsible for MNNG resistance in the drm1-1 strain,
additional information
-
construction of a dcd1DELTA enzyme-deficient strain, the deletion strain and the dcd1-S178F mutant strain are crossed with a rad52DELTA mgt1DELTA strain and haploid, double and triple mutant strains are generated, rad52DELTA mgt1DELTA dcd1-S178F and rad52DELTA mgt1DELTA dcd1DELTA strains show an increased level of MNNG resistance, while the only dcd1-deficient strain does not, overview, DCD1 deficiency in yeast results in elevation of dCTP pools and a slight decrease in dTTP pools and reduced the frequency of spontaneous and MNNGinduced G/C to A/T mutations, phenotypes, overview
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
after denaturation with EDTA, mutants R115E and R115Q restored 54% and 60% of original activities, wild-type enzyme only marginally restored; after treatment with guanidine-HCl, 6 M, complete reactivation on removal of denaturant by dilution or dialysis
-
after treatment with guanidine hydrochloride, 6 M, complete reactivation with 50 mM 2-mercaptoethanol, original hexameric structure restored
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
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Show AA Sequence (532 entries)
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