Cholesterol metabolism in the rat lactating mammary gland: the role of cholesteryl ester hydrolase.
An acid cholesteryl ester hydrolase activity associated with a fraction containing mitochondria and lysosomes from rat lactating mammary glands was found to have a pH optimum of 5.0. Its sedimentation pattern was closely related to that of the lysosomal enzyme markers acid phosphatase and beta-glucuronidase, suggesting that the activity is associated with the lysosomes. The enzyme was strongly inhibited by Cu2+, but was inhibited little by other divalent metal ions. Acid cholesteryl ester hydrolase activity was almost completely abolished by p-hydroxy-mercuribenzoate, but this effect was reversed in the presence of an equimolar concentration of reduced glutathione (GSH), indicating that the enzyme requires free sulfhydryl groups for activity. These properties are similar to those of acid, lysosomal cholesteryl ester hydrolases found in other tissues. Acid cholesteryl ester hydrolase activity was 8-14 fold higher in mammary tissue from lactating as compared to virgin rats. Neutral cholesteryl ester hydrolase activities associated with the microsomal and cytosolic subcellular fractions were also increased in lactating glands, but to a lesser extent. In addition, a 2-fold increase in the activities of both the acid and microsomal neutral enzymes was seen during the first few days of lactation, while the cytosolic neutral activity remained constant. These results suggest that mammary gland cholesteryl ester hydrolases have a role in the regulation of cholesterol metabolism in mammary cells, and in the provision of cholesterol for secretion into milk.
Botham
K M
KM
Department of Veterinary Basic Sciences, Royal Veterinary College, London, United Kingdom.
D000818:Animals; D015415:Biomarkers; D002413:Cations, Divalent / Q000494:pharmacology; D002729:Chloromercuribenzoates / Q000494:pharmacology; D002784:Cholesterol / Q000378:metabolism*; D003300:Copper / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D005260:Female; D006863:Hydrogen-Ion Concentration; D007774:Lactation / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology; D008321:Mammary Glands, Animal / Q000201:enzymology / Q000378:metabolism*; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D051381:Rats; D012969:Sodium Fluoride / Q000494:pharmacology; D002787:Sterol Esterase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013438:Sulfhydryl Compounds
D015415:Biomarkers; D002413:Cations, Divalent; D002729:Chloromercuribenzoates; D013438:Sulfhydryl Compounds; D003300:Copper; D012969:Sodium Fluoride; D002784:Cholesterol; D002787:Sterol Esterase