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Literature summary extracted from
- Characterization of a cytochrome P-450-dependent steroid hydroxylase system present in Bacillus megaterium (1976), J. Biol. Chem., 251, 2831-2838.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.15.8 | heme | the heme content of cytochrome P-450meg is 0.94 nmol of heme per nmol of cytochrome P-450 | Priestia megaterium |  |
| 1.14.15.8 | Iron | it is possible to resolve the hydroxylase system into three proteins: a strictly NADPH-dependent FMN-containing flavoprotein (megaredoxin reductase), an iron-sulfur protein (megaredoxin), and cytochrome P-450 (P-450meg). The activity of the 15beta-hydroxylase system is fully reconstituted upon combination of these three proteins and addition of NADPH. Megaredoxin has an apparent sulfur to iron ratio of 0.98 and shows g-signals at 1.90, 1.93, and 2.06 when analyzed by electron paramagnetic resonance spectroscopy | Priestia megaterium | |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.15.8 | 11beta-hydroxyprogesterone | - |
Priestia megaterium | |
| 1.14.15.8 | 16alpha-hydroxyprogesterone | - |
Priestia megaterium | |
| 1.14.15.8 | 17alpha,21-dihydroxyprogesterone | - |
Priestia megaterium | |
| 1.14.15.8 | 17alpha-hydroxyprogesterone | - |
Priestia megaterium | |
| 1.14.15.8 | 19-norandrostenedione | - |
Priestia megaterium | |
| 1.14.15.8 | 19-nortestosterone | - |
Priestia megaterium | |
| 1.14.15.8 | 20alpha-dihydroprogesterone | - |
Priestia megaterium | |
| 1.14.15.8 | deoxycorticosterone | - |
Priestia megaterium | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.15.8 | 0.06 | - |
deoxycorticosterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.095 | - |
corticosterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.143 | - |
progesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.45 | - |
20alpha-dihydroprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.56 | - |
17alpha-hydroxyprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.625 | - |
4-androstene-3,17-dione | 22°C | Priestia megaterium | |
| 1.14.15.8 | 1.11 | - |
testosterone | 22°C | Priestia megaterium | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.14.15.8 | 40000 | - |
sucrose density gradient centrifugation | Priestia megaterium |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.15.8 | Priestia megaterium | - |
ATCC 13368 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.15.8 | it is possible to resolve the hydroxylase system into three proteins: a strictly NADPH-dependent FMN-containing flavoprotein (megaredoxin reductase), an iron-sulfur protein (megaredoxin), and cytochrome P-450 (P-450meg). The activity of the 15beta-hydroxylase system is fully reconstituted upon combination of these three proteins and addition of NADPH. Megaredoxin has an apparent sulfur to iron ratio of 0.98 and shows g-signals at 1.90, 1.93, and 2.06 when analyzed by electron paramagnetic resonance spectroscopy | Priestia megaterium |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.15.8 | 11-deoxycorticosterone + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | 15beta-hydroxy-11-deoxycorticosterone + oxidized megaredoxin + H2O | - |
? | |
| 1.14.15.8 | 17alpha-hydroxyprogesterone + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | (15beta,17alpha)-15,17-dihydroxyprogesterone + oxidized megaredoxin + H2O | - |
? | |
| 1.14.15.8 | 20alpha-dihydroprogesterone + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | (15beta,20alpha)-15-hydroxy-20-dihydroprogesterone + oxidized megaredoxin + H2O | - |
? | |
| 1.14.15.8 | 4-androstene-3,17-dione + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | 15beta-hydroxyandrostene-3,17-dione + oxidized megaredoxin + H2O | - |
? | |
| 1.14.15.8 | corticosterone + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, preferred substrate, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | 15beta-hydroxycorticosterone + oxidized megaredoxin + H2O | - |
? | |
| 1.14.15.8 | additional information | hydroxylates a variety of 3-oxo-DELTA4-steroids in position 15beta in the presence of NADPH and O2. Ring A-reduced, aromatic, and 3beta-hydroxy-DELTA4-steroids do not serve as substrates | Priestia megaterium | ? | - |
? | |
| 1.14.15.8 | progesterone + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | 15beta-hydroxyprogesterone + oxidized megaredoxin + H2O | - |
? | |
| 1.14.15.8 | testosterone + reduced megaredoxin + O2 | megaredoxin is an iron-sulfur protein of Bacillus megaterium, megaredoxin has an apparent sulfur to iron ratio of 0.98 | Priestia megaterium | 15beta-hydroxytestosterone + oxidized megaredoxin + H2O | - |
? | |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.15.8 | cytochrome P-450meg | - |
Priestia megaterium |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.15.8 | 22 | - |
assay at | Priestia megaterium |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.15.8 | cytochrome P450meg | it is possible to resolve the hydroxylase system into three proteins: a strictly NADPH-dependent FMN-containing flavoprotein (megaredoxin reductase), an iron-sulfur protein (megaredoxin), and cytochrome P-450 (P-450meg). The activity of the 15beta-hydroxylase system is fully reconstituted upon combination of these three proteins and addition of NADPH. Megaredoxin has an apparent sulfur to iron ratio of 0.98 and shows g-signals at 1.90, 1.93, and 2.06 when analyzed by electron paramagnetic resonance spectroscopy | Priestia megaterium | |
| 1.14.15.8 | heme | the heme content of cytochrome P-450meg is 0.94 nmol of heme per nmol of cytochrome P-450 | Priestia megaterium | |
Ki Value [mM]
| EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.15.8 | 0.05 | - |
16alpha-hydroxyprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.05 | - |
20alpha-dihydroprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.055 | - |
17alpha-hydroxyprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.07 | - |
11beta-hydroxyprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.07 | - |
19-norandrostenedione | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.11 | - |
deoxycorticosterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.14 | - |
17alpha,21-dihydroxyprogesterone | 22°C | Priestia megaterium | |
| 1.14.15.8 | 0.16 | - |
19-nortestosterone | 22°C | Priestia megaterium | |
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