A functional cytochrome P450 lanosterol 14 alpha-demethylase CYP51 enzyme in the acrosome: transport through the Golgi and synthesis of meiosis-activating sterols.
Mammalian lanosterol 14 alpha-demethylase (CYP51) is a microsomal cytochrome P450 that demethylates lanosterol to FF-MAS, an oocyte meiosis-activating sterol and late intermediate of cholesterol biosynthesis. Herein we report CYP51 unequivocally localized to acrosomal membranes of male germ cells in mouse, bull, and ram, in which it synthesizes FF-MAS in the presence of the acrosomal form of nicotinamide adenine dinucleotide phosphate reduced-P450 reductase. In the mouse, CYP51 (53 kDa) resides in endoplasmic reticulum (ER) and Golgi during all phases of acrosome development, indicating an intracellular transport from ERs through the Golgi to the acrosome. CYP51 (50 kDa) also resides on acrosomal membranes of bull- and ram-ejaculated sperm. In mouse liver, a 53-kDa CYP51 is no longer detected in trans Golgi, suggesting retrieval back to the ER and no further transport to other organelles. Glycosylated high-molecular-mass CYP51-immunoreactive proteins in acrosomal membranes of bull and ram and Golgi-enriched fractions of mouse liver indicate that mammalian CYP51s are subjected to posttranslational modifications in the Golgi. In conclusion, CYP51 is the first cytochrome P450 enzyme to be detected on acrosomal membranes. It exhibits a unique, cell-type-specific intracellular transport that is in agreement with its cell-type-specific physiological role: production of cholesterol in the liver and sterols with signaling properties in sperm. Demethylation of lanosterol to FF-MAS by the acrosomal lanosterol 14 alpha-demethylase enzyme complex demonstrates for the first time the ability of ejaculate sperm to synthesize meiosis-activating sterols.
Cotman
M
M
Laboratory for Medical Genetics, Institute of Biochemistry, Faculty of Medicine, University of Ljubljana, SI-1000 Ljubljana, Slovenia.
Jezek
D
D
Fon Tacer
K
K
Frangez
R
R
Rozman
D
D
D000177:Acrosome / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D002458:Cell Fractionation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004542:Ejaculation; D006056:Golgi Apparatus / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D008297:Male; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D008808:Mice, Inbred CBA; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism*; D012756:Sheep; D013087:Spermatids / Q000201:enzymology; D013094:Spermatozoa / Q000201:enzymology; D058886:Sterol 14-Demethylase; D013261:Sterols / Q000096:biosynthesis; D013737:Testis / Q000166:cytology / Q000201:enzymology
D013261:Sterols; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; C548415:Cyp51 protein, mouse; D058886:Sterol 14-Demethylase; D009251:NADPH-Ferrihemoprotein Reductase