BRENDA - Enzyme Database show

Production, purification, and characterization of a Mg2+-responsive porphobilinogen synthase from pseudomonas aeruginosa

Frankenberg, N.; Heinz, D.W.; Jahn, D.; Biochemistry 38, 13968-13975 (1999)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
4.2.1.24
NH4+
stimulates
Pseudomonas aeruginosa
Cloned(Commentary)
EC Number
Commentary
Organism
4.2.1.24
expression in Escherichia coli
Pseudomonas aeruginosa
Crystallization (Commentary)
EC Number
Crystallization
Organism
4.2.1.24
hanging drop vapour diffusion method, crystals of the enzyme complex with levulinic acid solved at 1.67 A resolution, crystals belong to space group P42(1)2 with cell dimensions of a = b = 129.8 A, c = 86.7 A
Pseudomonas aeruginosa
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.24
2,2-difluorosuccinic acid
competitive
Pseudomonas aeruginosa
4.2.1.24
diethyl dicarbonate
-
Pseudomonas aeruginosa
4.2.1.24
EDTA
activity can be completely restored by addition of Mg2+ or Mn2+. Co2+, Zn2+, and Ni2+ partially restore EDTA-inhibited activity
Pseudomonas aeruginosa
4.2.1.24
levulinic acid
-
Pseudomonas aeruginosa
4.2.1.24
protoporphyrin IX
-
Pseudomonas aeruginosa
4.2.1.24
pyridoxal phosphate
-
Pseudomonas aeruginosa
4.2.1.24
succinic acid
noncompetitive
Pseudomonas aeruginosa
4.2.1.24
succinic acid monomethyl ester
competitive
Pseudomonas aeruginosa
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.24
0.33
-
5-aminolevulinate
pH 8.6 in Na-Hepes buffer
Pseudomonas aeruginosa
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.24
Co2+
partially restores activity after inhibition with EDTA
Pseudomonas aeruginosa
4.2.1.24
K+
stimulates
Pseudomonas aeruginosa
4.2.1.24
Li+
stimulates
Pseudomonas aeruginosa
4.2.1.24
Mg2+
can completrly restore activity after inhibition by EDTA; can completrly restore activity after inhibition by EDTA, stabilizes the oligomeric state but is not essential for octamer formation
Pseudomonas aeruginosa
4.2.1.24
Na+
stimulates
Pseudomonas aeruginosa
4.2.1.24
Ni2+
partially restores activity after inhibition with EDTA
Pseudomonas aeruginosa
4.2.1.24
Zn2+
partially restores activity after inhibition with EDTA
Pseudomonas aeruginosa
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.24
37832
-
6 * 37832, electrospray ionization mass spectrometry
Pseudomonas aeruginosa
4.2.1.24
280000
-
gel filtration
Pseudomonas aeruginosa
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.2.1.24
Pseudomonas aeruginosa
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.2.1.24
-
Pseudomonas aeruginosa
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.24
1
-
-
Pseudomonas aeruginosa
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.24
5-aminolevulinate + 5-aminolevulinate
-
664001
Pseudomonas aeruginosa
porphobilinogen + 2 H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.2.1.24
octamer
6 * 37832, electrospray ionization mass spectrometry
Pseudomonas aeruginosa
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.24
8.6
-
in presence of Mg2+ and K+
Pseudomonas aeruginosa
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
4.2.1.24
7.5
9.8
pH 7.5: about 35% of maximal activity, pH 9.8: about 50% of maximal activity
Pseudomonas aeruginosa
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
4.2.1.24
0.0025
-
protoporphyrin IX
-
Pseudomonas aeruginosa
4.2.1.24
0.03
-
diethyl dicarbonate
-
Pseudomonas aeruginosa
4.2.1.24
0.03
-
pyridoxal phosphate
-
Pseudomonas aeruginosa
4.2.1.24
1.5
-
levulinic acid
-
Pseudomonas aeruginosa
4.2.1.24
1.9
-
succinic acid monomethyl ester
-
Pseudomonas aeruginosa
4.2.1.24
7
-
2,2-difluorosuccinic acid
-
Pseudomonas aeruginosa
4.2.1.24
9
-
succinic acid
-
Pseudomonas aeruginosa
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
4.2.1.24
Pseudomonas aeruginosa
isoelectric focusing, pH-range 3-10
-
5.4
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
4.2.1.24
NH4+
stimulates
Pseudomonas aeruginosa
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.24
expression in Escherichia coli
Pseudomonas aeruginosa
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
4.2.1.24
hanging drop vapour diffusion method, crystals of the enzyme complex with levulinic acid solved at 1.67 A resolution, crystals belong to space group P42(1)2 with cell dimensions of a = b = 129.8 A, c = 86.7 A
Pseudomonas aeruginosa
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.24
2,2-difluorosuccinic acid
competitive
Pseudomonas aeruginosa
4.2.1.24
diethyl dicarbonate
-
Pseudomonas aeruginosa
4.2.1.24
EDTA
activity can be completely restored by addition of Mg2+ or Mn2+. Co2+, Zn2+, and Ni2+ partially restore EDTA-inhibited activity
Pseudomonas aeruginosa
4.2.1.24
levulinic acid
-
Pseudomonas aeruginosa
4.2.1.24
protoporphyrin IX
-
Pseudomonas aeruginosa
4.2.1.24
pyridoxal phosphate
-
Pseudomonas aeruginosa
4.2.1.24
succinic acid
noncompetitive
Pseudomonas aeruginosa
4.2.1.24
succinic acid monomethyl ester
competitive
Pseudomonas aeruginosa
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
4.2.1.24
0.0025
-
protoporphyrin IX
-
Pseudomonas aeruginosa
4.2.1.24
0.03
-
diethyl dicarbonate
-
Pseudomonas aeruginosa
4.2.1.24
0.03
-
pyridoxal phosphate
-
Pseudomonas aeruginosa
4.2.1.24
1.5
-
levulinic acid
-
Pseudomonas aeruginosa
4.2.1.24
1.9
-
succinic acid monomethyl ester
-
Pseudomonas aeruginosa
4.2.1.24
7
-
2,2-difluorosuccinic acid
-
Pseudomonas aeruginosa
4.2.1.24
9
-
succinic acid
-
Pseudomonas aeruginosa
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.24
0.33
-
5-aminolevulinate
pH 8.6 in Na-Hepes buffer
Pseudomonas aeruginosa
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.24
Co2+
partially restores activity after inhibition with EDTA
Pseudomonas aeruginosa
4.2.1.24
K+
stimulates
Pseudomonas aeruginosa
4.2.1.24
Li+
stimulates
Pseudomonas aeruginosa
4.2.1.24
Mg2+
can completrly restore activity after inhibition by EDTA; can completrly restore activity after inhibition by EDTA, stabilizes the oligomeric state but is not essential for octamer formation
Pseudomonas aeruginosa
4.2.1.24
Na+
stimulates
Pseudomonas aeruginosa
4.2.1.24
Ni2+
partially restores activity after inhibition with EDTA
Pseudomonas aeruginosa
4.2.1.24
Zn2+
partially restores activity after inhibition with EDTA
Pseudomonas aeruginosa
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.24
37832
-
6 * 37832, electrospray ionization mass spectrometry
Pseudomonas aeruginosa
4.2.1.24
280000
-
gel filtration
Pseudomonas aeruginosa
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.24
-
Pseudomonas aeruginosa
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.2.1.24
1
-
-
Pseudomonas aeruginosa
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.2.1.24
5-aminolevulinate + 5-aminolevulinate
-
664001
Pseudomonas aeruginosa
porphobilinogen + 2 H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.1.24
octamer
6 * 37832, electrospray ionization mass spectrometry
Pseudomonas aeruginosa
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.1.24
8.6
-
in presence of Mg2+ and K+
Pseudomonas aeruginosa
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
4.2.1.24
7.5
9.8
pH 7.5: about 35% of maximal activity, pH 9.8: about 50% of maximal activity
Pseudomonas aeruginosa
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
4.2.1.24
Pseudomonas aeruginosa
isoelectric focusing, pH-range 3-10
-
5.4