BRENDA - Enzyme Database

Characterization of recombinant human endothelial nitric-oxide synthase purified from the yeast Pichia pastoris

Leber, A.; Hemmens, B.; Klosch, B.; Goessler, W.; Raber, G.; Mayer, B.; Schmidt, K.; J. Biol. Chem. 274, 37658-37664 (1999)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.14.13.39
endothelial enzyme expressed in Pichia pastoris using a highly inducible alcohol oxidase promoter PAOX1
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.13.39
0.0039
-
L-arginine
recombinant from Pichia pastoris
Homo sapiens
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.13.39
Iron
0.8 mol per mol of subunit; heme-iron
Homo sapiens
1.14.13.39
Zinc
0.43 mol per mol of subunit
Homo sapiens
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.14.13.39
Homo sapiens
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.14.13.39
recombinant from Pichia pastoris
Homo sapiens
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.14.13.39
endothelium
-
Homo sapiens
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.14.13.39
0.19
-
purified recombinant enzyme
Homo sapiens
1.14.13.39
0.34
0.35
purified enzyme, recombinant, determined as NADPH-oxidase activity
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.13.39
2 L-arginine + 3 NADPH + 4 O2 + 3 H+
-
440216
Homo sapiens
2 L-citrulline + 2 NO + 3 NADP+ + 4 H2O
-
440216
Homo sapiens
?
1.14.13.39
oxidized cytochrome c + NADPH + O2
-
440216
Homo sapiens
reduced cytochrome c + NADP+ + H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.14.13.39
dimer
2 * 131000-133000, recombinant, SDS-PAGE
Homo sapiens
1.14.13.39
More
gel filtration: native protein is a dimer, especially in presence of tetrahydrobiopterin and L-arginine, but dissociates at low temperature to monomers during gel electrophoresis
Homo sapiens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.14.13.39
5,6,7,8-tetrahydro-L-biopterin
absolute requirement, recombinant from Pichia pastoris
Homo sapiens
1.14.13.39
Calmodulin
dependent on, endothelial enzyme
Homo sapiens
1.14.13.39
FAD
0.56 mol per mol of recombinant enzyme
Homo sapiens
1.14.13.39
FMN
0.79 mol per mol of recombinant enzyme
Homo sapiens
1.14.13.39
NADPH
-
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.13.39
endothelial enzyme expressed in Pichia pastoris using a highly inducible alcohol oxidase promoter PAOX1
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.14.13.39
5,6,7,8-tetrahydro-L-biopterin
absolute requirement, recombinant from Pichia pastoris
Homo sapiens
1.14.13.39
Calmodulin
dependent on, endothelial enzyme
Homo sapiens
1.14.13.39
FAD
0.56 mol per mol of recombinant enzyme
Homo sapiens
1.14.13.39
FMN
0.79 mol per mol of recombinant enzyme
Homo sapiens
1.14.13.39
NADPH
-
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.13.39
0.0039
-
L-arginine
recombinant from Pichia pastoris
Homo sapiens
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.14.13.39
Iron
0.8 mol per mol of subunit; heme-iron
Homo sapiens
1.14.13.39
Zinc
0.43 mol per mol of subunit
Homo sapiens
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.14.13.39
recombinant from Pichia pastoris
Homo sapiens
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.14.13.39
endothelium
-
Homo sapiens
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.14.13.39
0.19
-
purified recombinant enzyme
Homo sapiens
1.14.13.39
0.34
0.35
purified enzyme, recombinant, determined as NADPH-oxidase activity
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.14.13.39
2 L-arginine + 3 NADPH + 4 O2 + 3 H+
-
440216
Homo sapiens
2 L-citrulline + 2 NO + 3 NADP+ + 4 H2O
-
440216
Homo sapiens
?
1.14.13.39
oxidized cytochrome c + NADPH + O2
-
440216
Homo sapiens
reduced cytochrome c + NADP+ + H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.14.13.39
dimer
2 * 131000-133000, recombinant, SDS-PAGE
Homo sapiens
1.14.13.39
More
gel filtration: native protein is a dimer, especially in presence of tetrahydrobiopterin and L-arginine, but dissociates at low temperature to monomers during gel electrophoresis
Homo sapiens