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J. Biochem, 1967, Vol. 62, No. 3 345-352
© 1967 Japanese Biochemical Society

research-article

Extramitochondrial NADPH and Cholesterol Sidechain-cleavinfEnzyme System in the Heavy Mitochondria of Bovine Corpora Lutea

NAGASUGAMI YAGO, RALPH I. DORFMAN and ENRICO FORCHIELLI

From the Syntex Rtsiarch Center Stanford Industrial Park, Palo Alto, Calif., U.S.A.

1. The cholesterol sidechain-cleaving enzyme system in bovine corpora lutea was localized in the heavy mitochondria (the fraction sedimenting between 600xg and 3,500xg for 10 minutes). The enzyme system of the heavy mitochondria was tightly bound to membranous structures.

2. Exogenously added NADH and NADPH were not oxidized by the intact heavy mitochondria. The added NADPH was not accessible to the cholesterol sidechain-cleaving enzyme system of the intact heavy mitochondria.

3.The luteinizing hormone preparations (NIH-LH-B3 and -S8), respiratory inhibitors, uncouplers and the acetone powder preparation of the cellular soluble fraction were not effective in stimulating the utilization of external NADPH by the cholesterol sidechain-cleaving enzyme system of intact heavy mitochondria.

4.The external NADPH became accessible to the cholesterol sidechain-cleaving enzyme system of the heavy mitochondria after aging for a proper period.

5.These data suggested the existence of a permeability barrier to the extramitochondrial NADPH with regard to the cholesterol sidechain-cleaving enzyme system.

6.The significance of these findings was discussed in relation to the action mechanism of trophic hormones (ACTH and gonadotrophins).


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