BRENDA - Enzyme Database

A new subfamily of agmatinases present in methanogenic Archaea is Fe(II) dependent

Miller, D.; Xu, H.; White, R.H.; Biochemistry 51, 3067-3078 (2012)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
3.5.3.11
dithiothreitol
the enzyme requires the presence of dithiothreitol for maximum activity
Methanocaldococcus jannaschii
Cloned(Commentary)
EC Number
Commentary
Organism
3.5.3.11
overexpression in Escherichia coli
Methanocaldococcus jannaschii
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.5.3.11
C136A
mutant has 90% of the activity compared to wild type in the presence of Fe(II). As the wild-type enzyme the mutant enzyme requires dithiothreitol for activity
Methanocaldococcus jannaschii
3.5.3.11
C151S
mutant has 6% of the activity compared to wild type in the presence of Fe(II). When dithiothreitol is present in the reaction the mutant shows 24% of the wild type activity
Methanocaldococcus jannaschii
3.5.3.11
C229A
mutant has 92% of the activity compared to wild type in the presence of Fe(II)
Methanocaldococcus jannaschii
3.5.3.11
C71S
mutant has 4% of the activity compared to wild type in the presence of Fe(II). When dithiothreitol is present in the reaction the mutant shows 24% of the wild type activity
Methanocaldococcus jannaschii
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.3.11
iodoacetic acid
the enzyme is rendered inactive when the purified enzyme is incubated with dithiothreitol followed by excess iodoacetic acid
Methanocaldococcus jannaschii
3.5.3.11
L-arginine
44% inhibition at at 16.7 mM
Methanocaldococcus jannaschii
3.5.3.11
N-ethylmaleinimide
the enzyme is rendered inactive when the purified enzyme is incubated with dithiothreitol followed by excess N-ethylmaleimide
Methanocaldococcus jannaschii
3.5.3.11
putrescine
43% inhibition at 8.3 mM, 60% inhibition at 16.7 mM
Methanocaldococcus jannaschii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.3.11
2.4
-
agmatine
pH 7.5, 70C, wild-type enzyme
Methanocaldococcus jannaschii
3.5.3.11
5.8
-
agmatine
pH 7.5, 70C, mutant enzyme C229A
Methanocaldococcus jannaschii
3.5.3.11
6.2
-
agmatine
pH 7.5, 70C, mutant enzyme C151S
Methanocaldococcus jannaschii
3.5.3.11
9.2
-
agmatine
pH 7.5, 70C, mutant enzyme C136A
Methanocaldococcus jannaschii
3.5.3.11
11
-
agmatine
pH 7.5, 70C, mutant enzyme C71S
Methanocaldococcus jannaschii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.3.11
Fe2+
Fe(II)-dependent agmatinase. Requires the presence of 4 equivalents of Fe(II) for maximum activity
Methanocaldococcus jannaschii
3.5.3.11
Mn2+
supports activity at 15% of the activation by Fe2+
Methanocaldococcus jannaschii
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.3.11
40000
-
x * 40000, SDS-PAGE
Methanocaldococcus jannaschii
3.5.3.11
190000
-
gel filtration
Methanocaldococcus jannaschii
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.3.11
Methanocaldococcus jannaschii
Q57757
-
-
3.5.3.11
Methanocaldococcus jannaschii DSM 2661
Q57757
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.3.11
wild-type and mutant enzymes
Methanocaldococcus jannaschii
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
3.5.3.11
0.45
-
pH 7.5, 70C
Methanocaldococcus jannaschii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.3.11
agmatine + H2O
-
726980
Methanocaldococcus jannaschii
putrescine + urea
-
-
-
?
3.5.3.11
agmatine + H2O
-
726980
Methanocaldococcus jannaschii DSM 2661
putrescine + urea
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.5.3.11
?
x * 40000, SDS-PAGE
Methanocaldococcus jannaschii
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
3.5.3.11
70
-
assay at
Methanocaldococcus jannaschii
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.3.11
0.0075
-
agmatine
pH 7.5, 70C, mutant enzyme C71S
Methanocaldococcus jannaschii
3.5.3.11
0.0095
-
agmatine
pH 7.5, 70C, mutant enzyme C151S
Methanocaldococcus jannaschii
3.5.3.11
0.023
-
agmatine
pH 7.5, 70C, mutant enzyme C136A
Methanocaldococcus jannaschii
3.5.3.11
0.032
-
agmatine
pH 7.5, 70C, wild-type enzyme
Methanocaldococcus jannaschii
3.5.3.11
0.041
-
agmatine
pH 7.5, 70C, mutant enzyme C229A
Methanocaldococcus jannaschii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.3.11
7.5
-
assay at
Methanocaldococcus jannaschii
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.5.3.11
6.5
10
no activity is detected at pH 6.5 and the measured activity steadily increases to pH 10.0. Higher pH values are tested but the observation of activity is dependent on the buffer used and are not reported here. Agmatine shows no hydrolysis in the pH 10 and 11 buffers and assay conditions used
Methanocaldococcus jannaschii
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
3.5.3.11
dithiothreitol
the enzyme requires the presence of dithiothreitol for maximum activity
Methanocaldococcus jannaschii
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.3.11
overexpression in Escherichia coli
Methanocaldococcus jannaschii
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.5.3.11
C136A
mutant has 90% of the activity compared to wild type in the presence of Fe(II). As the wild-type enzyme the mutant enzyme requires dithiothreitol for activity
Methanocaldococcus jannaschii
3.5.3.11
C151S
mutant has 6% of the activity compared to wild type in the presence of Fe(II). When dithiothreitol is present in the reaction the mutant shows 24% of the wild type activity
Methanocaldococcus jannaschii
3.5.3.11
C229A
mutant has 92% of the activity compared to wild type in the presence of Fe(II)
Methanocaldococcus jannaschii
3.5.3.11
C71S
mutant has 4% of the activity compared to wild type in the presence of Fe(II). When dithiothreitol is present in the reaction the mutant shows 24% of the wild type activity
Methanocaldococcus jannaschii
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.3.11
iodoacetic acid
the enzyme is rendered inactive when the purified enzyme is incubated with dithiothreitol followed by excess iodoacetic acid
Methanocaldococcus jannaschii
3.5.3.11
L-arginine
44% inhibition at at 16.7 mM
Methanocaldococcus jannaschii
3.5.3.11
N-ethylmaleinimide
the enzyme is rendered inactive when the purified enzyme is incubated with dithiothreitol followed by excess N-ethylmaleimide
Methanocaldococcus jannaschii
3.5.3.11
putrescine
43% inhibition at 8.3 mM, 60% inhibition at 16.7 mM
Methanocaldococcus jannaschii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.3.11
2.4
-
agmatine
pH 7.5, 70C, wild-type enzyme
Methanocaldococcus jannaschii
3.5.3.11
5.8
-
agmatine
pH 7.5, 70C, mutant enzyme C229A
Methanocaldococcus jannaschii
3.5.3.11
6.2
-
agmatine
pH 7.5, 70C, mutant enzyme C151S
Methanocaldococcus jannaschii
3.5.3.11
9.2
-
agmatine
pH 7.5, 70C, mutant enzyme C136A
Methanocaldococcus jannaschii
3.5.3.11
11
-
agmatine
pH 7.5, 70C, mutant enzyme C71S
Methanocaldococcus jannaschii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.3.11
Fe2+
Fe(II)-dependent agmatinase. Requires the presence of 4 equivalents of Fe(II) for maximum activity
Methanocaldococcus jannaschii
3.5.3.11
Mn2+
supports activity at 15% of the activation by Fe2+
Methanocaldococcus jannaschii
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.3.11
40000
-
x * 40000, SDS-PAGE
Methanocaldococcus jannaschii
3.5.3.11
190000
-
gel filtration
Methanocaldococcus jannaschii
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.3.11
wild-type and mutant enzymes
Methanocaldococcus jannaschii
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
3.5.3.11
0.45
-
pH 7.5, 70C
Methanocaldococcus jannaschii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.3.11
agmatine + H2O
-
726980
Methanocaldococcus jannaschii
putrescine + urea
-
-
-
?
3.5.3.11
agmatine + H2O
-
726980
Methanocaldococcus jannaschii DSM 2661
putrescine + urea
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.5.3.11
?
x * 40000, SDS-PAGE
Methanocaldococcus jannaschii
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
3.5.3.11
70
-
assay at
Methanocaldococcus jannaschii
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.3.11
0.0075
-
agmatine
pH 7.5, 70C, mutant enzyme C71S
Methanocaldococcus jannaschii
3.5.3.11
0.0095
-
agmatine
pH 7.5, 70C, mutant enzyme C151S
Methanocaldococcus jannaschii
3.5.3.11
0.023
-
agmatine
pH 7.5, 70C, mutant enzyme C136A
Methanocaldococcus jannaschii
3.5.3.11
0.032
-
agmatine
pH 7.5, 70C, wild-type enzyme
Methanocaldococcus jannaschii
3.5.3.11
0.041
-
agmatine
pH 7.5, 70C, mutant enzyme C229A
Methanocaldococcus jannaschii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.3.11
7.5
-
assay at
Methanocaldococcus jannaschii
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.5.3.11
6.5
10
no activity is detected at pH 6.5 and the measured activity steadily increases to pH 10.0. Higher pH values are tested but the observation of activity is dependent on the buffer used and are not reported here. Agmatine shows no hydrolysis in the pH 10 and 11 buffers and assay conditions used
Methanocaldococcus jannaschii
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.5.3.11
0.00069
-
agmatine
pH 7.5, 70C, mutant enzyme C71S
Methanocaldococcus jannaschii
3.5.3.11
0.0015
-
agmatine
pH 7.5, 70C, mutant enzyme C151S
Methanocaldococcus jannaschii
3.5.3.11
0.0025
-
agmatine
pH 7.5, 70C, mutant enzyme C136A
Methanocaldococcus jannaschii
3.5.3.11
0.0077
-
agmatine
pH 7.5, 70C, mutant enzyme C229A
Methanocaldococcus jannaschii
3.5.3.11
0.013
-
agmatine
pH 7.5, 70C, wild-type enzyme
Methanocaldococcus jannaschii
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.5.3.11
0.00069
-
agmatine
pH 7.5, 70C, mutant enzyme C71S
Methanocaldococcus jannaschii
3.5.3.11
0.0015
-
agmatine
pH 7.5, 70C, mutant enzyme C151S
Methanocaldococcus jannaschii
3.5.3.11
0.0025
-
agmatine
pH 7.5, 70C, mutant enzyme C136A
Methanocaldococcus jannaschii
3.5.3.11
0.0077
-
agmatine
pH 7.5, 70C, mutant enzyme C229A
Methanocaldococcus jannaschii
3.5.3.11
0.013
-
agmatine
pH 7.5, 70C, wild-type enzyme
Methanocaldococcus jannaschii