Information on EC 3.5.3.18 - dimethylargininase

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

EC NUMBER
COMMENTARY hide
3.5.3.18
-
RECOMMENDED NAME
GeneOntology No.
dimethylargininase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Nomega,Nomega'-dimethyl-L-arginine + H2O = dimethylamine + L-citrulline
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
hydrolysis of linear amidines
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
Nomega,Nomega'-dimethyl-L-arginine dimethylamidohydrolase
Also acts on Nomega-methyl-L-arginine.
CAS REGISTRY NUMBER
COMMENTARY hide
123644-75-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
adult mongrels
-
-
Manually annotated by BRENDA team
White leg-horn chick embryos
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
dimethyl-L-arginine + H2O
L-citrulline + dimethylamine
show the reaction diagram
L-arginine + H2O
?
show the reaction diagram
-
-
-
-
?
N,N-dimethyl-L-arginine + H2O
dimethylamine + L-citrulline
show the reaction diagram
N-monomethyl-L-arginine + H2O
L-citrulline + methylamine
show the reaction diagram
N5-(methoxycarbamimidoyl)-L-ornithine + H2O
L-citrulline + ?
show the reaction diagram
-
assay at 37C, 30 min, pH 7.4
-
-
?
N5-(methylcarbamimidoyl)-L-ornithine + H2O
L-citrulline + ?
show the reaction diagram
-
assay at 37C, 30 min, pH 7.4
-
-
?
N5-[2,5-dihydro-1H-pyrrol-1-yl(imino)methyl]-L-ornithine + H2O
L-citrulline + ?
show the reaction diagram
-
assay at 37C, 30 min, pH 7.4
-
-
?
NG,NG-dimethyl-L-arginine + H2O
?
show the reaction diagram
NG,NG-dimethyl-L-arginine + H2O
dimethylamine + L-citrulline
show the reaction diagram
NG,NG-dimethyl-L-arginine + H2O
L-citrulline + dimethylamine
show the reaction diagram
NG,NG-dimethyl-L-homoarginine + H2O
?
show the reaction diagram
-
at 2.8% of the activity with NG,NG-dimethyl-L-arginine
-
-
?
NG-amino-L-arginine + H2O
?
show the reaction diagram
-
-
-
-
?
NG-hydroxy-L-arginine + H2O
?
show the reaction diagram
-
-
-
-
?
NG-methyl arginine + H2O
L-citrulline + methylamine
show the reaction diagram
-
-
-
-
?
NG-methyl-L-arginine + H2O
?
show the reaction diagram
-
-
-
-
?
NG-methyl-L-arginine + H2O
L-citrulline + methylamine
show the reaction diagram
NG-monomethyl-L-arginine + H2O
citrulline + methylamine
show the reaction diagram
Ngamma-monomethyl-L-arginine + H2O
L-citrulline + methylamine
show the reaction diagram
Nomega,Nomega'-dimethyl-L-arginine + H2O
dimethylamine + L-citrulline
show the reaction diagram
-
-
-
-
?
Nomega,Nomega-dimethyl-L-arginine + H2O
dimethylamine + L-citrulline
show the reaction diagram
-
-
-
?
Nomega-monomethyl-L-arginine + H2O
L-citrulline + methylamine
show the reaction diagram
-
-
-
?
S-methyl-L-thiocitrulline + H2O
?
show the reaction diagram
-
-
-
-
?
S-methyl-L-thiocitrulline + H2O
L-citrulline + methanethiol
show the reaction diagram
S-methyl-L-thiocitrulline + H2O
methanethiol + L-citrulline
show the reaction diagram
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
NG,NG-dimethyl-L-arginine + H2O
dimethylamine + L-citrulline
show the reaction diagram
NG,NG-dimethyl-L-arginine + H2O
L-citrulline + dimethylamine
show the reaction diagram
-
-
-
-
?
Nomega,Nomega'-dimethyl-L-arginine + H2O
dimethylamine + L-citrulline
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
DDAH1 forms a protein complex with Ras
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S)-2-amino-5-(ethanimidoylamino)pentanoic acid
-
-
(2S)-2-amino-5-(N'-butylcarbamimidamido)pentanoic acid
-
-
(2S)-2-amino-5-(pent-4-enimidoylamino)pentanoic acid
-
-
(2S)-2-amino-5-(pentanimidoylamino)pentanoic acid
-
-
(2S)-2-amino-5-(propanimidoylamino)pentanoic acid
-
-
(2S)-2-amino-5-[(4E)-hex-4-enimidoylamino]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(3,3,3-trifluoropropyl)carbamimidamido]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(3-methoxypropyl)carbamimidamido]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(4-amino-4-oxobutyl)carbamimidamido]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(but-3-en-1-yl)carbamimidamido]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(but-3-yn-1-yl)carbamimidamido]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(nitromethyl)carbamimidamido]pentanoic acid
-
-
(2S)-2-amino-5-[N'-(pent-4-en-1-yl)carbamimidamido]pentanoic acid
-
-
2,3-dimethoxy-1,4-naphthoquinone
-
1 mM
2-(N,N-diethylamino)-diazenolate-2-oxide
-
1 mM
2-(N,N-dimethylamino)-diazenolate-2-oxide
2-(N,N-dimethylamino)diazenolate-2-oxide
2-amino-4-(NG-methyl-guanidino)butanoic acid
2-chloroacetamidine
-
irreversible inhibition in a time- and concentration-dependent manner
2-hydroxymethyl-4-chloropyridine
-
solution studies support an inactivation mechanismin in which the active site Asp66 residue stabilizes the pyridinium form of the inactivator, which has enhanced reactivity toward the active site Cys, resulting in covalent bond formation, loss of the halide, and irreversible inactivation
4-hydroxy-2-nonenal
50 microM, 50% relative activity
4-hydroxy-nonenal
-
dose-dependently inhibits DDAH activity with 15% inhibition at 0.01 mM and complete inhibition at 0.5 mM
angiotensin II
1 microM, about 55% relative activity; 1 microM, about 55% relative activity
aprotinin inhibitor
-
NG,NG-dimethyl-L-arginine release from freeze-thaw whole blood 34% inhibited
-
CoCl2
-
-
cytokine induced nitric-oxide synthesis
-
-
D-arginine
-
0.1 mM D-arginine reduces DDAH activity to 24.7%
darbepoetin alpha
-
inhibits DDAH in a dose dependent manner, inhibition of DDAH is abolished by incubation with EPO antibody or with antioxidant pyrrolidine dithiocarbamate
-
epoetin beta
-
inhibits DDAH in a dose dependent manner, inhibition of DDAH is abolished by incubation with EPO antibody or with antioxidant pyrrolidine dithiocarbamate
-
erythropoetin
200 units/ml, about 75% relative activity; 200 units/ml, about 75% relative activity
-
glucose
glycosylated bovine serum albumin
;
-
H2O2
-
43% inhibition at 1 mM
homocysteine
hydrogen peroxide
100 microM, 50% relative activity
hyperglycemia
-
-
-
iodoacetamide
-
pH dependent DDAH inactivation, addition of 2.5 mM NG-methyl-L-arginine at pH 8.5 can prevent inactivation by idoacetamide, inactivation may be due to modification at the active site of DDAH, inactivation increases at higher pH values
l-257
-
-
L-arginine
L-citrulline
L-homocysteine
L-lysine
-
competitive inhibitor; potential competitive inhibitor
Leupeptin
-
NG,NG-dimethyl-L-arginine release from freeze-thaw whole blood 55% inhibited
lipopolysaccharide
1 microg/ml, 24 h, about 55% relative activity; 1 microg/ml, about 55% relative activity
low-density-lipoprotein
; 100 microg/ml
-
lysophosphatidylcholine
Mercuric chloride
;
methylamine
-
10 mM, 80% of activity
monocrotaline
-
DDAH activity and DDAH1, but not DDAH2, protein expression are significantly reduced after 60mg/kg monocrotaline treatment
N(G)-nitro-L-arginine methyl ester
-
reduced DDAH-2 mRNA level
N-(2-methoxyethyl)-L-arginine
complete inhibition at 100 microM; complete inhibition at 100 microM
N-(2-methoxyethyl)arginine
;
N-(2-methoxyethyl)arginine methyl esther
;
N-(but-3-yn-1-yl)-2-chloroethanimidamide
-
click chemistry mediated in vivo activity probe that labels the active fraction of DDAH-1 in intact mammalian cells and that can be blocked by the presence of competitive reversible and irreversible inhibitors
N-nitro-L-arginine methylester
N5-(1-iminoethyl)-L-ornithine
-
N5-(1-iminohexyl)-L-ornithine
-
N5-(1-iminopentyl)-L-ornithine
-
N5-(1-iminopropyl)-L-ornithine
crystallization data. Reversible competitive inhibition, consistent with a reversible covalent mode of DDAH inhibition by alkylamidine inhibitors
N5-(but-3-en-1-ylcarbamimidoyl)-L-ornithine
-
1 mM, 71% inhibition
N5-(nitrocarbamimidoyl)-L-ornithine
-
1 mM, 6% inhibition
N5-(prop-2-en-1-ylcarbamimidoyl)-L-ornithine
-
1 mM, 70% inhibition
N5-(prop-2-yn-1-ylcarbamimidoyl)-L-ornithine
-
1 mM, 83% inhibition
N5-(propylcarbamimidoyl)-L-ornithine
-
1 mM, 60% inhibition
N5-but-3-enimidoyl-L-ornithine
-
1 mM, 97% inhibition
N5-butanimidoyl-L-ornithine
-
1 mM, 78% inhibition
N5-ethanimidoyl-L-ornithine
-
1 mM, 24% inhibition
N5-propanimidoyl-L-ornithine
-
1 mM, 65% inhibition
N5-[(2,2,2-trifluoroethyl)carbamimidoyl]-L-ornithine
-
1 mM, 41% inhibition
N5-[(2-methoxyethyl)carbamimidoyl]-L-ornithine
-
1 mM, 89% inhibition
N5-[(3-amino-3-oxopropyl)carbamimidoyl]-L-ornithine
-
1 mM, 43% inhibition
N5-[(3E)-pent-3-enimidoyl]-L-ornithine
-
1 mM, 73% inhibition
N5-[imino(morpholin-4-yl)methyl]-L-ornithine
-
1 mM, 23% inhibition
Nicotine
50% decrease in DDAH-2 mRNA
nitric oxide
nitrite
-
1 mM
nitroglycerine
10 microM, 16 h, about 55% relative activity; 10 microM, about 55% relative activity
OH radical
-
47% inhibition at 1 mM
oxidized-low density lipoprotein
PD 404182
-
i.e. 6H-6-imino-(2,3,4,5-tetrahydropyrimido)[1,2-c]-[1,3]benzothiazine, potent active-site competitive inhibitor, about 75% inhibition at 0.05 mM
pentafluorophenyl sulfonates
-
between 30-76% DDAH inhibition depending on molecular structure of the pentafluoropenyl sulfonate
-
peroxynitrite
PMSF
-
NG,NG-dimethyl-L-arginine release from freeze-thaw whole blood 57% inhibited
protease inhibitor
-
NG,NG-dimethyl-L-arginine release from freeze-thaw whole blood 25-85% inhibited
-
S-2-amino-4(3-methylguanidino)butanoic acid
S-2-amino-4-[N-(2-methoxyethyl)guanidino]butanoic acid benzyl ester
;
S-nitroso-L-cysteine
-
9-15% inhibition of the zinc-free enzyme at 0.5 mM, S-nitrosylation of the active site Cys273 in DDAH-1
S-nitroso-L-homocysteine
sulfurosalicylic acid
-
inactivates DDAH
TNF-alpha
; 250 U/ml, 63% relative activity
-
Zinc acetate
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adiponectin
;
-
all-trans retinoic acid
-
inhibits cobalt chloride induced inhibition of DDAH
all-trans-retinoic acid
1.9fold increase in DDAH-2 mRNA and protein expression; 1.9fold increase in DDAH-2 mRNA and protein expression
aminoguanidine
;
D-histidine
-
activating ability in decreasing order: histamine, imidazole, L-histidine, D-histidine
epigallocatechin gallate
;
fenofibrate
;
histamine
-
activating ability in decreasing order: histamine, imidazole, L-histidine, D-histidine
imidazole
-
activating ability in decreasing order: histamine, imidazole, L-histidine, D-histidine
Insulin
;
-
Interleukin-1beta
2fold increase in DDAH-1 protein expression
-
L-histidine
-
activating ability in decreasing order: histamine, imidazole, L-histidine, D-histidine
nebivolol
2fold increase in DDAH-2 mRNA and protein expression
-
oestradiol
60-70% increase in DDAH-2 mRNA and protein expression
pioglitazone
2-3fold increase of DDAH-2 mRNA and protein levels
probucol
;
-
SNAP
-
increase of DDAH-2 mRNA and protein level, time and concentration dependent
-
taurin
taurine
telmisartan
visfatin
-
mRNA and protein expression of DDAH2 upregulated after 24 h stimulation of visfatin
-
vitamin E
xanthone
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.18
dimethyl-L-arginine
0.133
N,N-dimethyl-L-arginine
-
pH 7.4, 37C
-
0.13
N5-(methoxycarbamimidoyl)-L-ornithine
-
37C, 30 min, pH 7.4
0.106
N5-(methylcarbamimidoyl)-L-ornithine
-
37C, 30 min, pH 7.4
0.161
N5-[2,5-dihydro-1H-pyrrol-1-yl(imino)methyl]-L-ornithine
-
37C, 30 min, pH 7.4
0.039 - 9.24
NG,NG-dimethyl-L-arginine
33
NG,NG-Dimethyl-L-homoarginine
-
-
1.11
NG-Amino-L-arginine
-
-
2.3
NG-Hydroxy-L-arginine
-
-
0.044 - 0.67
NG-methyl-L-arginine
0.36 - 1.6
NG-monomethyl-L-arginine
0.11 - 13.7
Nomega,Nomega-dimethyl-L-arginine
0.0014 - 0.143
S-methyl-L-thiocitrulline
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.27 - 1.27
NG,NG-dimethyl-L-arginine
0.009
NG,NG-Dimethyl-L-homoarginine
Rattus norvegicus
-
-
0.12
NG-Amino-L-arginine
Pseudomonas aeruginosa
-
-
0.33
NG-Hydroxy-L-arginine
Pseudomonas aeruginosa
-
-
0.31 - 0.59
NG-methyl-L-arginine
0.094 - 0.34
NG-monomethyl-L-arginine
0.0015 - 0.0163
Nomega,Nomega-dimethyl-L-arginine
0.0097 - 1.3
S-methyl-L-thiocitrulline
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00012 - 0.127
Nomega,Nomega-dimethyl-L-arginine
1.58 - 13.55
S-methyl-L-thiocitrulline
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.155
(2S)-2-amino-5-(ethanimidoylamino)pentanoic acid
-
pH 7.4, 37C
0.09
(2S)-2-amino-5-(N'-butylcarbamimidamido)pentanoic acid
-
pH 7.4, 37C
0.002
(2S)-2-amino-5-(pent-4-enimidoylamino)pentanoic acid
-
pH 7.4, 37C
0.032
(2S)-2-amino-5-(pentanimidoylamino)pentanoic acid
-
pH 7.4, 37C
0.145
(2S)-2-amino-5-(propanimidoylamino)pentanoic acid
-
pH 7.4, 37C
0.036
(2S)-2-amino-5-[(4E)-hex-4-enimidoylamino]pentanoic acid
-
pH 7.4, 37C
0.606
(2S)-2-amino-5-[N'-(3,3,3-trifluoropropyl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C
0.013
(2S)-2-amino-5-[N'-(3-methoxypropyl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C, inhibitor is selective over NOS and over arginase
0.764
(2S)-2-amino-5-[N'-(4-amino-4-oxobutyl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C
0.058
(2S)-2-amino-5-[N'-(but-3-en-1-yl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C
0.017
(2S)-2-amino-5-[N'-(but-3-yn-1-yl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C
1.968
(2S)-2-amino-5-[N'-(nitromethyl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C
0.057
(2S)-2-amino-5-[N'-(pent-4-en-1-yl)carbamimidamido]pentanoic acid
-
pH 7.4, 37C
3.1
2-chloroacetamidine
-
-
0.4 - 4
L-lysine
0.057
N5-(but-3-en-1-ylcarbamimidoyl)-L-ornithine
-
-
0.058
N5-(prop-2-en-1-ylcarbamimidoyl)-L-ornithine
-
-
0.017
N5-(prop-2-yn-1-ylcarbamimidoyl)-L-ornithine
-
-
0.09
N5-(propylcarbamimidoyl)-L-ornithine
-
-
0.002
N5-but-3-enimidoyl-L-ornithine
-
-
0.032
N5-butanimidoyl-L-ornithine
-
-
0.145
N5-propanimidoyl-L-ornithine
-
-
0.013
N5-[(2-methoxyethyl)carbamimidoyl]-L-ornithine
-
-
0.036
N5-[(3E)-pent-3-enimidoyl]-L-ornithine
-
-
0.69
S-nitroso-L-homocysteine
-
in the presence of 5 mM EDTA, at pH 7.3 and 37C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05
4-hydroxy-nonenal
Homo sapiens
-
at 37C
0.131
L-arginine
Homo sapiens
O94760
pH 7.3, temperature not specified in the publication
3.7
L-citrulline
Homo sapiens
O94760
pH 7.3, temperature not specified in the publication
0.35
N-(but-3-yn-1-yl)-2-chloroethanimidamide
Homo sapiens
-
pH not specified in the publication, temperature not specified in the publication
0.99
N5-(1-iminoethyl)-L-ornithine
Homo sapiens
O94760
pH 7.3, temperature not specified in the publication
0.11
N5-(1-iminohexyl)-L-ornithine
Homo sapiens
O94760
pH 7.3, temperature not specified in the publication
0.0075
N5-(1-iminopentyl)-L-ornithine
Homo sapiens
O94760
pH 7.3, temperature not specified in the publication
0.052
N5-(1-iminopropyl)-L-ornithine
Homo sapiens
O94760
pH 7.3, temperature not specified in the publication
0.189
N5-(but-3-en-1-ylcarbamimidoyl)-L-ornithine
Homo sapiens
-
-
0.207
N5-(prop-2-en-1-ylcarbamimidoyl)-L-ornithine
Homo sapiens
-
-
0.055
N5-(prop-2-yn-1-ylcarbamimidoyl)-L-ornithine
Homo sapiens
-
-
0.283
N5-(propylcarbamimidoyl)-L-ornithine
Homo sapiens
-
-
0.013
N5-but-3-enimidoyl-L-ornithine
Homo sapiens
-
-
0.07
N5-butanimidoyl-L-ornithine
Homo sapiens
-
-
0.3
N5-propanimidoyl-L-ornithine
Homo sapiens
-
-
0.029
N5-[(2-methoxyethyl)carbamimidoyl]-L-ornithine
Homo sapiens
-
-
0.079
N5-[(3E)-pent-3-enimidoyl]-L-ornithine
Homo sapiens
-
-
0.009
PD 404182
Homo sapiens
-
at 37C, pH not specified in the publication
0.075
S-nitroso-L-homocysteine
Bos taurus
-
in 50 mM HEPES/NaOH (pH 7.3), 150 mM KCl, and 5 mM EDTA for 30 min at 37C
0.0088
Zinc acetate
Pseudomonas aeruginosa
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.55
-
37C
0.9
-
mice fed the high-methionine/low folate diet, heterozygous cystathionine beta-synthase
1.1
-
mice fed the high-methionine/low folate diet, homozygous cystathionine beta-synthase
1.6
-
mice fed the control diet
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.7
-
hydrolysis of NG-monomethyl-L-arginine
6.8
-
imidazole/HCl buffer
6.9
-
assay at
7.4
-
assay at
additional information
-
type of buffer dramatically influences enzymatic activity
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
both DDAHI and DDAHII proteins are detected in peri-infarct cardiomyocytes, while DDAHII immunoreactivity is additionally localized to infiltrating inflammatory cells and blood vessels in the healing infarct. Both plasma and left ventricular concentrations of the DDAH substrate, ADMA, are increased post-myocardial infarction
Manually annotated by BRENDA team
-
DDAH-2 protein expression is higher in osteoarthritic cartilage than in normal cartilage
Manually annotated by BRENDA team
-
7th cranial nerve
Manually annotated by BRENDA team
-
in healthy hearts, DDAHI is absent, and DDAHII is localized to endothelium and endocardium with a similar distribution to that of eNOS
Manually annotated by BRENDA team
-
DDAHII immunoreactivity is localized to infiltrating inflammatory cells and blood vessels in the healing infarct
Manually annotated by BRENDA team
-
normal human chondrocytes are isolated from knee cartilage obtained
Manually annotated by BRENDA team
-
both DDAH1 and 2 are expressed in the developing limb buds in patterns overlapping areas with high nitric oxide synthase activity
Manually annotated by BRENDA team
-
lowest activity in skeletal muscle
Manually annotated by BRENDA team
-
peritoneal
Manually annotated by BRENDA team
-
undifferentiated pheochromocytoma
Manually annotated by BRENDA team
-
endothelial cells
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
conventional immunofluorescence and confocal microscopy reveal the presence of DDAH-2 in the mitochondria of IL-1beta-stimulated chondrocytes. Blocking the translocation to mitochondria reduces the NO production induced by IL-1beta
Manually annotated by BRENDA team
-
cardiac myocytes
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
29000
-
monomer, hydrodynamic measurement
30420
-
mass spectrometry, mutant H162G
30560
-
fully inactivated wild-type DDAH by 2-chloroacetamidine
31200
-
electrospray ionization mass spectrometry
32000
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gel filtration
33440
-
electrospray ionization mass spectrometry
34000
-
DDAH1, SDS-PAGE
37000
-
DDAH-1, SDS-PAGE
58000
-
dimer, hydrodynamic measurement
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
2 * 29000, dynamic equilibrium between monomer and homodimer, hydrodynamic measurements
homodimer
-
PaDDAH exists in dynamic equilibrium between symmetric homodimer and monomer states
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by sitting drop vapor diffusion, at 1.08-2.0 A resolution and at pH 6.3 and pH 9.0
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N5-(1-iminopropyl)-L-ornithine:DDAH-1 complex, to 1.9 A resolution, covalent bond formation between the inhibitors amidino carbon and the active-site Cys274, and solution studies show reversible competitive inhibition, consistent with a reversible covalent mode of DDAH inhibition by alkylamidine inhibitors
A 2.18 A resolution X-ray crystal structure of the inactivated complex elucidates the orientation of the inactivator and its covalent attachment to the active site Cys
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8.5
-
stable between
288971
10
-
activity decreases at pH values above 10
687644
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
2 h, holo-enzyme retains ca. 56% of the original activity, no residual activity of the apoenzyme. Thermal denaturation of both protein forms is irreversible
additional information
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-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Zn2+ has a structure-stabilizing role
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-30C, 0.1 M sodium phosphate buffer, pH 7.0, 10% glycerol, 1 mM 2-mercaptoethanol
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
anion exchange chromatography and gel filtration chromatography
-
by immobilized nickel-affinity chromatography and gel filtration
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HisTrap column chromatography and HiTrap Q Sepharose column chromatography
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Ni-NTA affinity resin chromatography and phenyl-Sepharose column chromatography
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recombinant enzyme, partial
to more than 95% homogeneity
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using Ni-NTA-agarose
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wild type to more than 98% homogeneity, by Ni-NTA affinity chromatography, mutant required additional purification step, ion-exchange chromatography followed by affinity chromatography, mutant purified to more than 95% homogeneity
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloned into a pET-28a expression vector, N-terminal His6-tagged protein overexpressed in Escherichia coli BL21 (DE3)
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expressed in Escherichia coli BL21 (DE3) cells
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expressed in Escherichia coli BL21star cells
-
expressed in Mus musculus
-
expression in Escherichia coli
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expression in Escherichia coli BL21
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expression in Escherichia coli BL21 (DE3)
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expression in sEnd.1 cells
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human DDAH-1 is expressed in Escherichia coli BL21
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isoform DDAH-1 bearing an N-terminal Myc-tag, expression in HEK 293T cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
alpha-lipoic acid increases the gene expression and activity of DDAH, and signal transducer and activator of transcription STAT3 phosphorylation. Transfection of STAT3 increases DDAH II promoter activity, and alpha-lipoic acid amplifies it. alpha-Lipoic acid-induced increase in DDAH II promoter activity is attenuated in the promoter that has a mutation in the putative STAT3-binding site
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DDAH isoforms are expressed in a dynamic,overlapping pattern during embryonic development
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DDAH-2 protein expression is higher in osteoarthritic cartilage than in normal cartilage; in chondrocyte mitochondria extracts, DDAH-2 expression is significantly increased after exposure to IL-1beta
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following myocardial infarction, left ventricular DDAH activity is increased to 210% of control
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treatment of mice with farnesoid X receptor agonist 3-(2,6-dichlorophenyl)-4-(3'-carboxy-2-chlorostilben-4-yl)oxymethyl-5-isopropylisoxazole, i.e. GW4064, leads to increased expression of DDAH-1 and cationic amino acid transporter CAT-1 in both liver and kidney
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C275A
-
retains a kcat value about half of wild type protein
H173A
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no detectable activity
D244N
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wild type and D244N DDAH variants both form covalent adducts upon incubation with 2-hydroxymethyl-4-chloropyridine, showing mass additions that are consistent with covalent attachment of one equivalent of hydroxymethylpyridine to each enzyme. These results also indicate that Asp244 is not essential for covalent modification to occur
D66N
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the C249S mutant and D66N mutant are both incapable of forming a covalent adduct when incubated with 2-hydroxymethyl-4-chloropyridine
E33H
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weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
E33Q
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weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
N36D
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expressed in Escherichia coli at levels similar to the wild type
N36H
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weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
N36W
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more stable homodimer than the wild type protein, expressed in Escherichia coli at levels similar to the wild type
Q43H
-
weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
Q43R
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weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
R40E
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weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
R40E-R98H
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exclusively monomeric, expressed in Escherichia coli at levels similar to the wild type
R40E/R98H
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about 95% of wild type activity, stable monomer
R40W
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not expressed in Escherichia coli at levels similar to the wild type
R98H
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weakening of the homodimer interaction observed for wild type protein, expressed in Escherichia coli at levels similar to the wild type
R98N
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not expressed in Escherichia coli at levels similar to the wild type
S248N
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mutant is still capable of substrate turnover and is still inactivated by 2-hydroxymethyl-4-chloropyridine with a second order inactivation rate constant that is approximately 2fold less than wild type DDAH
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
inhibition by 2-(N,N-dimethylamino)-diazenolate-2-oxide is reversed by dithiothreitol
method for reversible preparation of metal-free and metal-containing enzyme
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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use of inhibitor N-(but-3-yn-1-yl)-2-chloroethanimidamide as a broad-specificity probe for labeling endogenous DDAH isoforms and enzymes with similar pharmacophores. Inhibitor labels the active fraction of DDAH-1 in intact mammalian cells and can be blocked by the presence of competitive reversible and irreversible inhibitors. Incorporation of the alkyne tag allows to derivatize with a variety of reagents after in vivo tagging
medicine
additional information
Show AA Sequence (263 entries)
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