Cloned (Comment) | Organism |
---|---|
expression of the N-terminal His-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
5 mg/ml purified recombinant wild-type and selenomethionine-enzyme, sitting drop vapour diffusion method, 4°C, equal volumes of protein solution, containing 10 mM Tris-HCl, pH 7.5, 1 mM dithiothreitol, and reservoir solution, containing 20% 2-methyl-2,4-pentanediol, 100 mM MES, pH 6.0, X-ray diffraction structure determination and analysis at 1.85-2.1 A resolution | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
A66V | clinical mutation, altered tertiary structure | Homo sapiens |
E127A | site-directed mutagenesis, unaltered activity level compared to the wild-type enzyme | Homo sapiens |
E249stop | clinical mutation, truncated protein, loss of helix 12 | Homo sapiens |
G225S | clinical mutation, altered tertiary structure | Homo sapiens |
I129T | clinical mutation, altered tertiary structure | Homo sapiens |
K220A | site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme | Homo sapiens |
L4F | clinical mutation, altered tertiary structure | Homo sapiens |
additional information | the naturally occuring clinical mutations lead to loss in activity due to structural alterations in the enzymes, overview | Homo sapiens |
P248Q | clinical mutation, altered tertiary structure | Homo sapiens |
P53L | clinical mutation, altered tertiary structure | Homo sapiens |
R65A | site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme | Homo sapiens |
S197A | site-directed mutagenesis, unaltered activity level compared to the wild-type enzyme | Homo sapiens |
S212P | clinical mutation, altered tertiary structure | Homo sapiens |
S63A | site-directed mutagenesis, unaltered activity level compared to the wild-type enzyme | Homo sapiens |
T103A | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Homo sapiens |
T227A | site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme | Homo sapiens |
T228A | site-directed mutagenesis, conserved residue near to the active site cleft, reduced activity compared to the wild-type enzyme | Homo sapiens |
T228M | clinical mutation, altered tertiary structure | Homo sapiens |
T62A | site-directed mutagenesis, conserved residue near to the active site cleft, unaltered activity level compared to the wild-type enzyme | Homo sapiens |
V3F | clinical mutation, altered tertiary structure | Homo sapiens |
Y168A | site-directed mutagenesis, reduced activity compared to the wild-type enzyme | Homo sapiens |
Y19C | clinical mutation, altered tertiary structure | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
hydroxymethylbilane | Homo sapiens | linear tetrapyrrole, fourth step in the biosynthesis of porphyrin, essential reaction, decreased enzyme activity leads to the autosomal recessive disorder congenital erythropetic porphyria | uroporphyrinogen-III + H2O | macrocyclic uroporphyrinogen III is a precurosr for synthesis of diverse compounds, e.g. heme, siroheme, chlorophyll, F430, and vitamin B12 | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P10746 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
hydroxymethylbilane = uroporphyrinogen III + H2O | in the presence of hydroxymethylbilane synthase EC 4.3.1.8, enzyme forms uroporphyrinogen III from porphobilinogen, reaction mechanism, does not require acid/base catalysis, active site structure | Homo sapiens |
Renatured (Comment) | Organism |
---|---|
recombinant His-tagged wild-type enzyme from Escherichia coli, to homogeneity | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
hydroxymethylbilane | - |
Homo sapiens | uroporphyrinogen-III + H2O | - |
? | |
hydroxymethylbilane | linear tetrapyrrole, fourth step in the biosynthesis of porphyrin, essential reaction, decreased enzyme activity leads to the autosomal recessive disorder congenital erythropetic porphyria | Homo sapiens | uroporphyrinogen-III + H2O | macrocyclic uroporphyrinogen III is a precurosr for synthesis of diverse compounds, e.g. heme, siroheme, chlorophyll, F430, and vitamin B12 | ? |
Subunits | Comment | Organism |
---|---|---|
More | observed interdomain flexibility might be important for catalysis, the active site is located between the domains | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
U3S | - |
Homo sapiens |
Uroporphyrinogen III synthase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.2 | - |
assay at | Homo sapiens |