BRENDA - Enzyme Database

Identification and characterization of plant agmatine iminohydrolase, the last missing link in polyamine biosynthesis of plants

Janowitz, T.; Kneifel, H.; Piotrowski, M.; FEBS Lett. 544, 258-261 (2003)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.3.12
DNA and amino acid sequence determination and analysis, expression as C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli
Arabidopsis thaliana
Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.5.3.12
C180A
site-directed mutagenesis, conserved catalytically relevant Cys, 35% of wild-type activity, inhibition with iodoacetamide is easier abolished by substrate agmatine
Arabidopsis thaliana
3.5.3.12
C366A
site-directed mutagenesis, conserved catalytically relevant Cys, 15% of wild-type activity, inhibition with iodoacetamide is easier abolished by substrate agmatine
Arabidopsis thaliana
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.3.12
iodoacetamide
about 70% inhibition at 2 mM, wild-type enzyme
Arabidopsis thaliana
3.5.3.12
NEM
1 mM
Arabidopsis thaliana
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.3.12
43000
-
2 * 43000, recombinant His-tagged wild-type enzyme, SDS-PAGE
Arabidopsis thaliana
3.5.3.12
81000
-
recombinant His-tagged wild-type enzyme, gel filtration
Arabidopsis thaliana
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.3.12
Arabidopsis thaliana
Q8GWW7
gene At5g08170
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.3.12
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by Ni2+-chelate affinity chromatography
Arabidopsis thaliana
Reaction
EC Number
Reaction
Commentary
Organism
3.5.3.12
agmatine + H2O = N-carbamoylputrescine + NH3
the plant enzyme also catalyzes the reactions of EC 2.1.3.3, ornithine carbamoyltransferase, EC 2.1.3.6, putrescine carbamoyltransferase, and EC 2.7.2.2, carbamate kinase, thus functioning as a putrescine synthase, converting agmatine and ornithine into putrescine and citrulline, respectively, Cys180 and Cys366 are involved in catalysis but are not essential, residues are located in the substrate binding pocket
Arabidopsis thaliana
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.3.12
26.4
-
purified recombinant wild-type enzyme
Arabidopsis thaliana
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.3.12
agmatine + H2O
highly specific for
655571
Arabidopsis thaliana
N-carbamoylputrescine + NH3
-
-
-
?
3.5.3.12
additional information
substrate specificity, no activity with guanidinoacetic acid, beta-guanidinopropionic acid, gamma-guanidinobutyric acid, L-arginine, L-citrulline, L-glutamine, L-ornithine, N-carbamoyl-beta-alanine, N-carbamoyl-D,L-aspartic acid, putrescine, spermidine, allylcyanide, beta-cyano-L-alanine, indole-3-acetonitrile, 3-phenylpropionitrile
655571
Arabidopsis thaliana
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
3.5.3.12
dimer
2 * 43000, recombinant His-tagged wild-type enzyme, SDS-PAGE
Arabidopsis thaliana
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.5.3.12
35
40
-
Arabidopsis thaliana
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.3.12
7
-
-
Arabidopsis thaliana
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.3.12
DNA and amino acid sequence determination and analysis, expression as C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli
Arabidopsis thaliana
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.5.3.12
C180A
site-directed mutagenesis, conserved catalytically relevant Cys, 35% of wild-type activity, inhibition with iodoacetamide is easier abolished by substrate agmatine
Arabidopsis thaliana
3.5.3.12
C366A
site-directed mutagenesis, conserved catalytically relevant Cys, 15% of wild-type activity, inhibition with iodoacetamide is easier abolished by substrate agmatine
Arabidopsis thaliana
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.5.3.12
iodoacetamide
about 70% inhibition at 2 mM, wild-type enzyme
Arabidopsis thaliana
3.5.3.12
NEM
1 mM
Arabidopsis thaliana
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.3.12
43000
-
2 * 43000, recombinant His-tagged wild-type enzyme, SDS-PAGE
Arabidopsis thaliana
3.5.3.12
81000
-
recombinant His-tagged wild-type enzyme, gel filtration
Arabidopsis thaliana
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.3.12
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by Ni2+-chelate affinity chromatography
Arabidopsis thaliana
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.3.12
26.4
-
purified recombinant wild-type enzyme
Arabidopsis thaliana
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.3.12
agmatine + H2O
highly specific for
655571
Arabidopsis thaliana
N-carbamoylputrescine + NH3
-
-
-
?
3.5.3.12
additional information
substrate specificity, no activity with guanidinoacetic acid, beta-guanidinopropionic acid, gamma-guanidinobutyric acid, L-arginine, L-citrulline, L-glutamine, L-ornithine, N-carbamoyl-beta-alanine, N-carbamoyl-D,L-aspartic acid, putrescine, spermidine, allylcyanide, beta-cyano-L-alanine, indole-3-acetonitrile, 3-phenylpropionitrile
655571
Arabidopsis thaliana
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.5.3.12
dimer
2 * 43000, recombinant His-tagged wild-type enzyme, SDS-PAGE
Arabidopsis thaliana
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.5.3.12
35
40
-
Arabidopsis thaliana
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.5.3.12
7
-
-
Arabidopsis thaliana