BRENDA - Enzyme Database

Mutational analysis of substrate recognition by human arginase type I -agmatinase activity of the N130D variant

Alarcon, R.; Orellana, M.S.; Neira, B.; Uribe, E.; Garcia, J.R.; Carvajal, N.; FEBS J. 273, 5625-5631 (2006)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.3.1
expressed in Escherichia coli strain JM109
Homo sapiens
3.5.3.11
expressed in Escherichia coli strain JM109
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.5.3.1
N130D
17% activity compared to the wild type enzyme, the mutant is also active on agmatine, the N130D mutant enzyme is inactive after dialysis against EDTA and assay in the absence of Mn2+
Homo sapiens
3.5.3.1
N130Q
after dialysis against EDTA and assay in the absence of Mn2+, the wild type-like N130Q mutant enzyme exhibits 50% activity
Homo sapiens
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.3.1
EDTA
after dialysis against EDTA and assay in the absence of Mn2+, the wild type enzyme exhibits 50% activity
Homo sapiens
3.5.3.1
Guanidinium chloride
-
Homo sapiens
3.5.3.1
L-ornithine
-
Homo sapiens
3.5.3.11
putrescine
linear competitive inhibitor
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.3.1
1.5
-
L-arginine
wild type enzyme, at 37°C
Homo sapiens
3.5.3.1
13.3
-
L-arginine
mutant enzyme N130D, at 37°C
Homo sapiens
3.5.3.11
1.4
-
agmatine
-
Homo sapiens
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.3.1
Mn2+
essentiell for activity
Homo sapiens
3.5.3.11
Mn2+
totally dependent on Mn2+ for catalytic activity
Homo sapiens
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.3.1
120000
-
SDS-PAGE
Homo sapiens
3.5.3.11
120000
-
SDS-PAGE
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.3.1
L-arginine + H2O
Homo sapiens
-
L-ornithine + urea
-
-
?
3.5.3.1
additional information
Homo sapiens
the wild type enzyme does not hydrolyze 1-amino-4-guanidinobutane (agmatine)
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.3.1
Homo sapiens
-
-
-
3.5.3.11
Homo sapiens
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.3.1
DEAE-cellulose column chromatography
Homo sapiens
3.5.3.11
DEAE-cellulose column chromatography
Homo sapiens
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.5.3.11
liver
-
Homo sapiens
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.3.1
L-arginine + H2O
-
686646
Homo sapiens
L-ornithine + urea
-
-
-
?
3.5.3.1
additional information
the wild type enzyme does not hydrolyze 1-amino-4-guanidinobutane (agmatine)
686646
Homo sapiens
?
-
-
-
-
3.5.3.11
agmatine + H2O
the N130D mutant of arginase type I is able to cleave not only arginine but also agmatine (1-amino-4-guanidinobutane)
686646
Homo sapiens
putrescine + urea
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.3.1
33
-
L-arginine
mutant enzyme N130D, at 37°C
Homo sapiens
3.5.3.1
190
-
L-arginine
wild type enzyme, at 37°C
Homo sapiens
3.5.3.11
3
-
agmatine
-
Homo sapiens
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.5.3.1
1
-
L-ornithine
wild type enzyme
Homo sapiens
3.5.3.1
7.8
-
L-ornithine
mutant enzyme N130D
Homo sapiens
3.5.3.1
39
-
Guanidinium chloride
mutant enzyme N130D
Homo sapiens
3.5.3.1
56
-
Guanidinium chloride
wild type enzyme
Homo sapiens
3.5.3.11
1.3
-
putrescine
-
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.3.1
expressed in Escherichia coli strain JM109
Homo sapiens
3.5.3.11
expressed in Escherichia coli strain JM109
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.5.3.1
N130D
17% activity compared to the wild type enzyme, the mutant is also active on agmatine, the N130D mutant enzyme is inactive after dialysis against EDTA and assay in the absence of Mn2+
Homo sapiens
3.5.3.1
N130Q
after dialysis against EDTA and assay in the absence of Mn2+, the wild type-like N130Q mutant enzyme exhibits 50% activity
Homo sapiens
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.3.1
EDTA
after dialysis against EDTA and assay in the absence of Mn2+, the wild type enzyme exhibits 50% activity
Homo sapiens
3.5.3.1
Guanidinium chloride
-
Homo sapiens
3.5.3.1
L-ornithine
-
Homo sapiens
3.5.3.11
putrescine
linear competitive inhibitor
Homo sapiens
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.5.3.1
1
-
L-ornithine
wild type enzyme
Homo sapiens
3.5.3.1
7.8
-
L-ornithine
mutant enzyme N130D
Homo sapiens
3.5.3.1
39
-
Guanidinium chloride
mutant enzyme N130D
Homo sapiens
3.5.3.1
56
-
Guanidinium chloride
wild type enzyme
Homo sapiens
3.5.3.11
1.3
-
putrescine
-
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.3.1
1.5
-
L-arginine
wild type enzyme, at 37°C
Homo sapiens
3.5.3.1
13.3
-
L-arginine
mutant enzyme N130D, at 37°C
Homo sapiens
3.5.3.11
1.4
-
agmatine
-
Homo sapiens
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.3.1
Mn2+
essentiell for activity
Homo sapiens
3.5.3.11
Mn2+
totally dependent on Mn2+ for catalytic activity
Homo sapiens
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.3.1
120000
-
SDS-PAGE
Homo sapiens
3.5.3.11
120000
-
SDS-PAGE
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.5.3.1
L-arginine + H2O
Homo sapiens
-
L-ornithine + urea
-
-
?
3.5.3.1
additional information
Homo sapiens
the wild type enzyme does not hydrolyze 1-amino-4-guanidinobutane (agmatine)
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.3.1
DEAE-cellulose column chromatography
Homo sapiens
3.5.3.11
DEAE-cellulose column chromatography
Homo sapiens
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.5.3.11
liver
-
Homo sapiens
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.3.1
L-arginine + H2O
-
686646
Homo sapiens
L-ornithine + urea
-
-
-
?
3.5.3.1
additional information
the wild type enzyme does not hydrolyze 1-amino-4-guanidinobutane (agmatine)
686646
Homo sapiens
?
-
-
-
-
3.5.3.11
agmatine + H2O
the N130D mutant of arginase type I is able to cleave not only arginine but also agmatine (1-amino-4-guanidinobutane)
686646
Homo sapiens
putrescine + urea
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.3.1
33
-
L-arginine
mutant enzyme N130D, at 37°C
Homo sapiens
3.5.3.1
190
-
L-arginine
wild type enzyme, at 37°C
Homo sapiens
3.5.3.11
3
-
agmatine
-
Homo sapiens