Separation of two forms of cytochrome P450 adrenal cortex mitochondria

Authors:
  • RHR. Hume
    R. Hume
  • GBG.S. Boyd
    G.S. Boyd
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Abstract
The separation of two distinct forms of cytochrome P450 from adrenal cortex mitochondria has been achieved by the following steps; (1) lyophilisation (2) iso‐octane extraction, (3) (NH4)2SO4 fractionation in the presence of sodium cholate. The fraction precipitating between 25–35 percent (NH4)2SO4 gave a difference spectrum with 11‐deoxycorticosterone (11‐DOC) but not with 20α‐hydroxycholesterol (20α‐HOC). This fraction showed high 11β‐hydroxylase activity but low activity for side chain cleavage of cholesterol (S.C.C.). The fraction precipitating between 45–60 percent (NH4)2SO4 gave a difference spectrum with 20α‐HOC but not with 11‐DOC and exhibited high S.C.C. activity but low 11β‐hydroxylase activity. The absorption spectrum of the 45–60 percent fraction indicated a preponderance of high spin hemoprotein (λmax 395 nm).

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