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Plastid associated porphobilinogen synthase from toxoplasma gondii: kinetic and structural properties validate therapeutic potential

Shanmugam, D.; Wu, B.; Ramirez, U.; Jaffe, E.K.; Roos, D.S.; J. Biol. Chem. 285, 22122-22131 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.2.1.24
recombinant TgPBGS is expressed in Escherichia coli as a His-tagged fusion protein
Toxoplasma gondii
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.24
DELTA646-658
a mutant enzyme lacking the C-terminal 13 amino acids distinguishing parasite PBGS from plant and animal enzymes is purified as a dimer, suggesting that the C-terminus is required for octamer stabilisation
Toxoplasma gondii
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.24
succinylacetone
-
Toxoplasma gondii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.24
0.15
-
5-aminolevulinic acid
-
Toxoplasma gondii
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
4.2.1.24
apicoplast
-
Toxoplasma gondii
20011
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.24
Mg2+
enzyme responds to Mg2+ but not to Zn2+, enzyme shows two Mg2+ affinities
Toxoplasma gondii
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.24
40000
-
recombinant TgPBGS also shows low levels of dimers, 2 * 40000 Da, SDS-PAGE; recombinant TgPBGS is purified as a stable octamer, 8 * 40000 Da, SDS-PAGE
Toxoplasma gondii
4.2.1.24
320000
-
gel filtration, recombinant TgPBGS is purified as a stable octamer
Toxoplasma gondii
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.2.1.24
Toxoplasma gondii
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.2.1.24
using Ni-NTA chromatography
Toxoplasma gondii
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
0.4
-
-
Toxoplasma gondii
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-aminolevulinic acid + 5-aminolevulinic acid
-
704678
Toxoplasma gondii
porphobilinogen + 2 H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.2.1.24
dimer
recombinant TgPBGS also shows low levels of dimers, 2 * 40000 Da, SDS-PAGE
Toxoplasma gondii
4.2.1.24
octamer
recombinant TgPBGS is purified as a stable octamer, 8 * 40000 Da, SDS-PAGE
Toxoplasma gondii
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.1.24
37
-
assay at
Toxoplasma gondii
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
4.2.1.24
7
8.5
-
Toxoplasma gondii
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
4.2.1.24
0.001
-
-
Toxoplasma gondii
succinylacetone
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.24
recombinant TgPBGS is expressed in Escherichia coli as a His-tagged fusion protein
Toxoplasma gondii
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.2.1.24
DELTA646-658
a mutant enzyme lacking the C-terminal 13 amino acids distinguishing parasite PBGS from plant and animal enzymes is purified as a dimer, suggesting that the C-terminus is required for octamer stabilisation
Toxoplasma gondii
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
4.2.1.24
0.001
-
-
Toxoplasma gondii
succinylacetone
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.2.1.24
succinylacetone
-
Toxoplasma gondii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.1.24
0.15
-
5-aminolevulinic acid
-
Toxoplasma gondii
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
4.2.1.24
apicoplast
-
Toxoplasma gondii
20011
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.1.24
Mg2+
enzyme responds to Mg2+ but not to Zn2+, enzyme shows two Mg2+ affinities
Toxoplasma gondii
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.24
40000
-
recombinant TgPBGS also shows low levels of dimers, 2 * 40000 Da, SDS-PAGE; recombinant TgPBGS is purified as a stable octamer, 8 * 40000 Da, SDS-PAGE
Toxoplasma gondii
4.2.1.24
320000
-
gel filtration, recombinant TgPBGS is purified as a stable octamer
Toxoplasma gondii
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.24
using Ni-NTA chromatography
Toxoplasma gondii
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
0.4
-
-
Toxoplasma gondii
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-aminolevulinic acid + 5-aminolevulinic acid
-
704678
Toxoplasma gondii
porphobilinogen + 2 H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.1.24
dimer
recombinant TgPBGS also shows low levels of dimers, 2 * 40000 Da, SDS-PAGE
Toxoplasma gondii
4.2.1.24
octamer
recombinant TgPBGS is purified as a stable octamer, 8 * 40000 Da, SDS-PAGE
Toxoplasma gondii
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.1.24
37
-
assay at
Toxoplasma gondii
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
4.2.1.24
7
8.5
-
Toxoplasma gondii