[Reconstitution of the cholesterol-hydroxylating system from adrenal cortex mitochondria by cytochrome P-450scc, adrenodoxin and the redox-mediator neutral red].
It has been found that 3-amino-7-dimethylamino-2-methylphenazine (neutral red, NR) is responsible for the electron transport from the cathode to adrenodoxin (Ad) and cytochrome P-450 (P-450scc) from adrenal cortex mitochondria inaccessible to direct electrochemical reduction under native conditions. The rate constant for Ad reduction by this mediator is equal to 1.1 x 10(5) M-1 s-1 at 25 degrees C; the values of enthalpy and entropy for the activation reaction are 26.6 kJ.mol-1 and -59.6 J.mol-1 deg-1, respectively. Using the shunted electron transport chain NR----Ad----P-450scc, the cholesterol conversion into pregnenolone in an electrochemical cell was performed. Pregnenolone was found to be the sole steroid product of this reaction. Superoxide dismutase and catalase had no effect on the activity of the shunted system. After removal or substitution of Ad the apo-Ad hemoprotein was reduced in a non-productive manner. Under identical reconstitution conditions methylviologen was ineffective as an electron carrier.
Gur'ev
O L
OL
Gilevich
S N
SN
Chashchin
V L
VL
D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000325:Adrenodoxin / Q000378:metabolism*; D000818:Animals; D002374:Catalase / Q000378:metabolism; D002417:Cattle; D002784:Cholesterol / Q000378:metabolism*; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism*; D004579:Electron Transport / Q000187:drug effects; D006900:Hydroxylation; D066298:In Vitro Techniques; D008928:Mitochondria / Q000378:metabolism; D009499:Neutral Red / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011284:Pregnenolone / Q000096:biosynthesis; D013482:Superoxide Dismutase / Q000378:metabolism; D013816:Thermodynamics
D000325:Adrenodoxin; D009499:Neutral Red; D011284:Pregnenolone; D002784:Cholesterol; D002374:Catalase; D002786:Cholesterol Side-Chain Cleavage Enzyme; D013482:Superoxide Dismutase
Rekonstruktsiia kholesteringidroksiliruiushcheĭ sistemy iz mitokhondriĭ kory nadpochechnikov na osnove tsitokhroma P-450scc, adrenodoxina i redoks-mediatora neĭtral'nogo krasnogo.