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 | 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 | Comment | Organism | Structure |
---|---|---|---|
O2 | - |
Streptomyces sp. |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces sp. | P12676 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Streptomyces sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cholesterol + O2 | - |
Streptomyces sp. | cholest-4-en-3-one + H2O2 | - |
? |
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 maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
1.381 | - |
O2 | mutant enzyme F359W | Streptomyces sp. | |
1.383 | - |
O2 | wild-type enzyme | Streptomyces sp. |