Cloned (Comment) | Organism |
---|---|
Escherichia coli BL21(DE3) pLysS with pUHD2 plasmid | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
vapor diffusion method, hanging drops with 20 mM Tris-HCl buffer, pH 8.2, containing 5 mM dithiothreitol and reservoir solution consisting of 0.6 M ammonium sulfate, 1.2 M lithium sulfate, 5% ethylene glycol, 50 mM sodium citrate, pH 5.6, and 50 mM dithiothreitol, diffraction data are collected at -173°C in 30% glycerol cryoprotected protein crystals | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
R167Q | 8.5% activity, reduced pH optimum from 8.0 to 6.0, accumulates long-lived intermediate complexes | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4 porphobilinogen + H2O | Homo sapiens | - |
hydroxymethylbilane + 4 NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P08397 | - |
- |
Purification (Comment) | Organism |
---|---|
cells centrifuged, washed with 0.9 M NaCl, re-centrifuged, washed with 20 mM Tris-HCl buffer, pH 8.2, with 5 mM dithiothreitol, and 200 microM PMSF, sonication, heating to 60°C under N2 gas, cooled to 4°C, supernatant applied to a Pharmacia 50 K/30 column packed with DEAE-Sephacel anion-exchange resin, equilibrated with 50 mM Tris-HCl buffer, pH 8.2 containing 5 mM dithiothreitol, and 100 microM PMSF, elution with 0-70 mM KCl gradient, active fractions are pooled and concentrated by ultrafiltration with a PM-10 membrane, further concentrated with a Centricon YM-10 centrifugal filter and gel-filtered on a Hiload 16/60 Superdex G-75 column, equilibrated with 100 mM Tris-HCl buffer, pH 8.2, containing 5 mM dithiothreitol and 100 microM PMSF, concentrated with a Centricon YM-10, buffer-exchanged into 20 mM Tris-HCl buffer, pH 8.2, containing 5 mM dithiothreitol with a Pharmacia PD10 column | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.002 | - |
R167Q mutant, pH 8.0 | Homo sapiens |
0.023 | - |
wild type, pH 8.0 | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4 porphobilinogen + H2O | - |
Homo sapiens | hydroxymethylbilane + 4 NH3 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
human PBGD | - |
Homo sapiens |
human porphobilinogen deaminase | - |
Homo sapiens |
hydroxymethylbilane synthase | - |
Homo sapiens |
preuroporphyrinogen synthase | - |
Homo sapiens |
uPBGD | ubiquitous PBGD, espressed in all cells | Homo sapiens |
uroporphyrinogen I synthase | - |
Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
dipyrromethane | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | mutation causes acute intermittent porphyria | Homo sapiens |
metabolism | third enzyme in heme biosynthesis pathway in mammals | Homo sapiens |