Modulation of epididymal delta 4-steroid 5 alpha-reductase activity in vitro by the phospholipid environment.
Epididymal 5 alpha-reductase converts testosterone to 5 alpha-dihydrotestosterone. The enzyme is localized to the nuclear and microsomal membranes, and using two approaches, we investigated the relationship between 5 alpha-reductase activity and the membrane environment. In the first, nuclear and microsomal membrane fractions were treated with phospholipases to modify specifically the structure of the phospholipid component of the membranes, and the effects of these treatments on the kinetic parameters of 5 alpha-reductase were examined. The second approach was to observe the effects of phospholipids of known structure on solubilized 5 alpha-reductase activity. Treatment of the membrane fractions with phospholipase C increased the Km(app) of both the nuclear and microsomal 5 alpha-reductases for testosterone. Phospholipase A2 treatment also increased the Km(app) of the microsomal enzyme, but in contrast, the Km(app) of the nuclear 5 alpha-reductase for testosterone was unaffected. This demonstrated a fundamental difference in the role of the membrane environment in the expression of 5 alpha-reductase activity in these subcellular compartments. The ability of phospholipids to enhance the activity of solubilized 5 alpha-reductase was highly specific and structure related. Only phosphatidylcholines containing either unsaturated acyl chains or saturated acyl chains of 12 carbon atoms were found to activate 5 alpha-reductase. The most potent activator was dilauroyl phosphatidylcholine, which reduced the Km(app) values of both nuclear and microsomal 5 alpha-reductases for testosterone, without affecting the concentration of active 5 alpha-reductase (Vmax(app) ). This is the first time that an activator of 5 alpha-reductase has been found. These findings suggest that epididymal 5 alpha-reductase activity may be regulated by changes in the phospholipid environment.
D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D004822:Epididymis / Q000201:enzymology*; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D008563:Membrane Lipids / Q000502:physiology*; D008861:Microsomes / Q000201:enzymology*; D009685:Nuclear Envelope / Q000201:enzymology*; D010088:Oxidoreductases / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000494:pharmacology; D010741:Phospholipases A / Q000494:pharmacology; D054467:Phospholipases A2; D010743:Phospholipids / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains; D010738:Type C Phospholipases / Q000494:pharmacology
D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; C015153:1,2-dilauroylphosphatidylcholine; D010088:Oxidoreductases; D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases