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Literature summary for 1.6.2.2 extracted from

  • Davis, C.A.; Crowley, L.J.; Barber, M.J.
    Cytochrome b5 reductase: the roles of the recessive congenital methemoglobinemia mutants P144L, L148P, and R159* (2004), Arch. Biochem. Biophys., 431, 233-244.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)-RIL Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
L148P 31% of wild type activity, reduced temperature stability and resistance against limited proteolysis with trypsin, increased affinity for NAD+ Rattus norvegicus
P144L 28% of wild type activity, reduced temperature stability and resistance against limited proteolysis with trypsin, increased affinity for NAD+ Rattus norvegicus
P144L/L148P 8% of wild type activity, reduced temperature stability and resistance against limited proteolysis with trypsin, increased affinity for NAD+ Rattus norvegicus
R159 deletion mutant, could not be successfully expressed Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
NAD+ competitive, stronger inhibition of mutant enzymes compared to wild type enzyme Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.008
-
ferricyanide L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
0.008
-
ferricyanide P144L mutant enzyme, pH 7.0, 25°C Rattus norvegicus
0.008
-
ferricyanide P144L/L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
0.008
-
ferricyanide wild type enzyme, pH 7.0, 25°C Rattus norvegicus
0.01
-
ferricytochome b5 L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
0.012
-
ferricytochome b5 P144L mutant enzyme, pH 7.0, 25°C Rattus norvegicus
0.013
-
ferricytochome b5 P144L/L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
0.013
-
ferricytochome b5 wild type enzyme, pH 7.0, 25°C Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane full length form contains a 3 kDa membrane anchoring domain Rattus norvegicus 16020
-
soluble truncated form lacking the membrane anchoring domain, present in erythrocytes Rattus norvegicus
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
SDS-PAGE Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 ferricytochrome b5 + NADH Rattus norvegicus involved in the synthesis of fatty acids and cholesterol, and in the oxidation of xenobiotics, enzyme defects causes methemoglobinemia type I or type II 2 ferrocytochrome b5 + NAD+ + H+
-
?

Organism

Organism UniProt Comment Textmining
Rattus norvegicus P20070
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzymes Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ferricyanide + NADH
-
Rattus norvegicus 2 ferrocyanide + NAD+ + H+
-
?
2 ferricytochrome b5 + NADH
-
Rattus norvegicus 2 ferrocytochrome b5 + NAD+ + H+
-
?
2 ferricytochrome b5 + NADH involved in the synthesis of fatty acids and cholesterol, and in the oxidation of xenobiotics, enzyme defects causes methemoglobinemia type I or type II Rattus norvegicus 2 ferrocytochrome b5 + NAD+ + H+
-
?
NADH + ferricytochrome b5
-
Rattus norvegicus NAD+ + H+ + ferrocytochrome b5
-
r
NADH + ferricytochrome b5
-
Rattus norvegicus NAD+ + H+ + 2 ferrocytochrome b5
-
r

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
33
-
ferricytochome b5 P144L/L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
100
-
ferricyanide P144L/L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
117
-
ferricytochome b5 P144L mutant enzyme, pH 7.0, 25°C Rattus norvegicus
130
-
ferricytochome b5 L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
283
-
ferricyanide P144L mutant enzyme, pH 7.0, 25°C Rattus norvegicus
300
-
ferricyanide L148P mutant enzyme, pH 7.0, 25°C Rattus norvegicus
417
-
ferricytochome b5 wild type enzyme, pH 7.0, 25°C Rattus norvegicus
800
-
ferricyanide wild type enzyme, pH 7.0, 25°C Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
FAD non-covalentely bound prosthetic group Rattus norvegicus
NADH
-
Rattus norvegicus