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Literature summary for 1.3.3.4 extracted from

  • Maneli, M.H.; Corrigall, A.V.; Klump, H.H.; Davids, L.M.; Kirsch, R.E.; Meissner, P.N.
    Kinetic and physical characterisation of recombinant wild-type and mutant human protoporphyrinogen oxidases (2003), Biochim. Biophys. Acta, 1650, 10-21.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain JM109 Homo sapiens

Protein Variants

Protein Variants Comment Organism
G11A site-directed mutagenesis, 0.02% activity compared to the wild-type enzyme Homo sapiens
G14A site-directed mutagenesis, 42.6% activity compared to the wild-type enzyme Homo sapiens
G9A site-directed mutagenesis, 0.52% activity compared to the wild-type enzyme Homo sapiens
H20P site-directed mutagenesis, 0.16% activity compared to the wild-type enzyme Homo sapiens
additional information identification of several mutations of the enzyme encoding gene, responsible for variegate porphyria, overview Homo sapiens
R168C site-directed mutagenesis, 17.5% activity compared to the wild-type enzyme Homo sapiens
R59I site-directed mutagenesis, 1.5% activity compared to the wild-type enzyme Homo sapiens
R59K site-directed mutagenesis, 37.6% activity compared to the wild-type enzyme Homo sapiens
R59S site-directed mutagenesis, 2.6% activity compared to the wild-type enzyme Homo sapiens
R59W site-directed mutagenesis, 0.28% activity compared to the wild-type enzyme, highly reduced FAD binding Homo sapiens
Y348C site-directed mutagenesis, 8.6% activity compared to the wild-type enzyme Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
acifluorfen inhibition potency with recombinant wild-type and mutant enzymes, overview Homo sapiens
bilirubin inhibition potency with recombinant wild-type and mutant enzymes, overview Homo sapiens
biliverdin IX hydrochloride inhibition potency with recombinant wild-type and mutant enzymes, overview Homo sapiens
methyl acifluorfen inhibition potency with recombinant wild-type and mutant enzymes, overview Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00083
-
protoporphyrinogen-IX recombinant mutant R59K, pH 8.2 Homo sapiens
0.00085
-
protoporphyrinogen-IX recombinant wild-type enzyme and mutant G14A, pH 8.2 Homo sapiens
0.001
-
protoporphyrinogen-IX recombinant mutant R168C, pH 8.2 Homo sapiens
0.00107
-
protoporphyrinogen-IX recombinant mutant Y348, pH 7.8 Homo sapiens
0.00126
-
protoporphyrinogen-IX recombinant mutant R59W, pH 8.2 Homo sapiens
0.0017
-
protoporphyrinogen-IX recombinant mutant R59S, pH 8.2 Homo sapiens
0.00209
-
protoporphyrinogen-IX recombinant mutant R59I, pH 8.2 Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrial inner membrane
-
Homo sapiens 5743
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
51000
-
x * 51000, SDS-PAGE Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
protoporphyrinogen-IX + 3 O2 Homo sapiens penultimate enzyme in the heme biosynthesis, several mutations of the enzyme encoding gene are responsible for variegate porphyria protoporphyrin-IX + 3 H2O2
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain JM109, to homogeneity Homo sapiens

Renatured (Commentary)

Renatured (Comment) Organism
renaturation and refolding after thermal denaturation and unfolding at 75°C by cooling to 15°C, pH 7.2 Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Homo sapiens
0.62
-
recombinant mutant Y348C Homo sapiens
1.26
-
recombinant mutant R168C Homo sapiens
2.69
-
recombinant mutant R59K Homo sapiens
3.04
-
recombinant mutant G14A Homo sapiens
7.15
-
recombinant wild-type enzyme Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
protoporphyrinogen-IX + 3 O2 penultimate enzyme in the heme biosynthesis, several mutations of the enzyme encoding gene are responsible for variegate porphyria Homo sapiens protoporphyrin-IX + 3 H2O2
-
?
protoporphyrinogen-IX + 3 O2 roles of 59 arginine residues and glycine residues in the flavin binding site in catalysis and cofactor binding Homo sapiens protoporphyrin-IX + 3 H2O2
-
?

Subunits

Subunits Comment Organism
? x * 51000, SDS-PAGE Homo sapiens
More secondary structure analysis, wild-type and mutant enzymes, structure/function relationship mutant R59W Homo sapiens

Synonyms

Synonyms Comment Organism
PPOX
-
Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
recombinant wild-type and mutants, stable at Homo sapiens
47.6
-
50% inactivation of mutant R168C Homo sapiens
50.2
-
50% inactivation of mutant Y348C Homo sapiens
53
-
50% inactivation of mutant R59W Homo sapiens
54.2
-
50% inactivation of mutant R59I Homo sapiens
56.3
-
50% inactivation of mutant R59K Homo sapiens
56.8
-
50% inactivation of recombinant wild-type enzyme Homo sapiens
57
-
50% inactivation of mutant G14A Homo sapiens
60.3
-
50% inactivation of mutant R59S Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.04
-
protoporphyrinogen-IX recombinant mutant R59W, pH 8.2 Homo sapiens
0.15
-
protoporphyrinogen-IX recombinant mutant R59I, pH 8.2 Homo sapiens
0.17
-
protoporphyrinogen-IX recombinant mutant R59S, pH 8.2 Homo sapiens
0.53
-
protoporphyrinogen-IX recombinant mutant Y348, pH 7.8 Homo sapiens
1.02
-
protoporphyrinogen-IX recombinant mutant R168C, pH 8.2 Homo sapiens
2.57
-
protoporphyrinogen-IX recombinant mutant R59K, pH 8.2 Homo sapiens
3.9
-
protoporphyrinogen-IX recombinant mutant G14A, pH 8.2 Homo sapiens
5.95
-
protoporphyrinogen-IX recombinant wild-type enzyme, pH 8.2 Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
mutant Y348C Homo sapiens
8.2
-
except for mutant Y348C Homo sapiens

Cofactor

Cofactor Comment Organism Structure
FAD required, binding site, structure, and mechanism, roles of 59 arginine residues and glycine residues in the flavin binding site in catalysis and cofactor binding Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.00011
-
acifluorfen recombinant mutant R59I, pH 8.2 Homo sapiens
0.00018
-
acifluorfen recombinant mutant R59K, pH 8.2 Homo sapiens
0.00023
-
acifluorfen recombinant wild-type, pH 8.2 Homo sapiens
0.00027
-
acifluorfen recombinant mutant R59S, pH 8.2 Homo sapiens
0.00031
-
acifluorfen recombinant mutant G14A, pH 8.2 Homo sapiens
0.00033
-
acifluorfen recombinant mutant R59W, pH 8.2 Homo sapiens
0.003
-
acifluorfen recombinant mutant R168C, pH 8.2 Homo sapiens
0.00472
-
acifluorfen recombinant mutant Y348C, pH 7.8 Homo sapiens