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Literature summary for 1.16.3.1 extracted from

  • Vashchenko, G.; Macgillivray, R.T.
    Functional role of the putative iron ligands in the ferroxidase activity of recombinant human hephaestin (2012), J. Biol. Inorg. Chem., 17, 1187-1195.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in expressed in baby hamster kidney cells Homo sapiens

Protein Variants

Protein Variants Comment Organism
D616A/H621A/E960A/H965A iron binding sites are mutated but one is unaffected. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. kcat (Fe2+) up to 10fold decreased compared to wild-type. Mutant does not retain a high-affinity iron oxidation component Homo sapiens
E264A/H269A/D616A/H621A iron binding sites are mutated but one is unaffected. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. Km for high-affinity oxidation of Fe2+ decreased compared to wild-type. kcat (Fe2+) up to 10fold decreased compared to wild-type. Only mutant that retains a high-affinity iron oxidation component Homo sapiens
E264A/H269A/D616A/H621A/E960A/H965A all three iron binding sites are mutated. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. kcat (Fe2+) up to 75 fold decreased compared to wild-type. Mutant does not retain a high-affinity iron oxidation component Homo sapiens
E264A/H269A/E960A/H965A iron binding sites are mutated but one is unaffected. Km and kcat for substrate 4-phenylenediamine decreased compared to wild-type. kcat (Fe2+) up to 10fold decreased compared to wild-type. Mutant does not retain a high-affinity iron oxidation component Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0015
-
4-phenylenediamine pH 5, 20°C Homo sapiens
0.0035
-
Fe2+ pH 5, 20°C, Km for high-affinity iron-binding sites using a two-component Eadie-Hofstee plot, Vmax: 67 nmol/min Homo sapiens
0.004
-
Fe2+ pH 5, 20°C Homo sapiens
0.0107
-
Fe2+ pH 5, 20°C, Km for low-affinity iron-binding sites using a two-component Eadie-Hofstee plot, Vmax: 286 nmol/min Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ rhHp possesses both high-affinity and low-affinity binding sites for ferrous iron Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-phenylenediamine + Fe2+ + O2
-
Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
hephaestin
-
Homo sapiens
rhHp
-
Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
Fe2+ pH 5, 20°C Homo sapiens
0.48
-
4-phenylenediamine pH 5, 20°C Homo sapiens