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Literature summary for 1.1.3.6 extracted from

  • Chen, L.; Lyubimov, A.Y.; Brammer, L.; Vrielink, A.; Sampson, N.S.
    The binding and release of oxygen and hydrogen peroxide are directed by a hydrophobic tunnel in cholesterol oxidase (2008), Biochemistry, 47, 5368-5377.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.3.6

Crystallization (Commentary)

Crystallization (Comment) Organism
structure of the F359W mutant enzyme, hanging drop vapor diffusion method. In the atomic-resolution structure of F359W, the indole ring of the tryptophan completely fills the tunnel and is observed in only a single conformation Streptomyces sp.

Protein Variants

Protein Variants Comment Organism
F359W kcat for the F359W mutant-catalyzed reaction decreases 13fold relative to that of the wild-type-catalyzed reaction. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for the mutant enzyme. Kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutant, but not with the wild-type enzyme. In the atomic-resolution structure of F359W, the indole ring of the tryptophan completely fills the tunnel and is observed in only a single conformation. The size of the indole is proposed to limit conformational rearrangement of residue 359 that leads to tunnel opening in the wild-type enzyme Streptomyces sp.
G347N mutant can not be saturated with oxygen. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for the mutant enzyme. Kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutant, but not with the wild-type enzyme Streptomyces sp.
N485D mutant can not be saturated with oxygen. Transfer of hydride from the sterol to the flavin prosthetic group is no longer rate-limiting for the mutant enzyme. Kinetic cooperativity with respect to molecular oxygen is observed with the tunnel mutant, but not with the wild-type enzyme Streptomyces sp.

Inhibitors

Inhibitors Comment Organism Structure
O2
-
 Streptomyces sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0015
-
 cholesterol mutant enzyme G347N, determined by cholest-4-en-3-one detection Streptomyces sp.
0.0027
-
 cholesterol mutant enzyme F359W, determined by cholest-4-en-3-one detection Streptomyces sp.
0.0027
-
 cholesterol wild-type enzyme, determined by cholest-4-en-3-one detection Streptomyces sp.
0.0027
-
 cholesterol wild-type enzyme, determined by H2O2 detection Streptomyces sp.
0.004
-
 cholesterol mutant enzyme F359W, determined by H2O2 detection Streptomyces sp.
0.005
-
 cholesterol mutant enzyme G347N, determined by H2O2 detection Streptomyces sp.
0.0062
-
 cholesterol mutant enzyme N485D, determined by H2O2 detection Streptomyces sp.
0.007
-
 cholesterol mutant enzyme N485D, determined by cholest-4-en-3-one detection Streptomyces sp.
0.3
-
 O2 wild-type enzyme Streptomyces sp.
0.617
-
 O2 mutant enzyme F359W Streptomyces sp.

Organism

Organism UniProt Comment Textmining
Streptomyces sp. P12676
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Streptomyces sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cholesterol + O2
-
 Streptomyces sp. cholest-4-en-3-one + H2O2
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.035
-
 cholesterol mutant enzyme N485D, determined by H2O2 detection Streptomyces sp.
0.073
-
 cholesterol mutant enzyme N485D, determined by cholest-4-en-3-one detection Streptomyces sp.
0.85
-
 cholesterol mutant enzyme G347N, determined by H2O2 detection Streptomyces sp.
0.86
-
 cholesterol mutant enzyme F359W, determined by cholest-4-en-3-one detection Streptomyces sp.
1
-
 cholesterol mutant enzyme G347N, determined by cholest-4-en-3-one detection Streptomyces sp.
1.3
-
 cholesterol mutant enzyme F359W, determined by H2O2 detection Streptomyces sp.
42
-
 cholesterol wild-type enzyme, determined by cholest-4-en-3-one detection Streptomyces sp.
47
-
 cholesterol wild-type enzyme, determined by H2O2 detection Streptomyces sp.

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.381
-
 O2 mutant enzyme F359W Streptomyces sp.
1.383
-
 O2 wild-type enzyme Streptomyces sp.