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Abstract 1. 1. Subcellular fractions were obtained bydifferential centrifugation of the homogenate of spheroplasts prepared from aerobically cultured Saccharomyces cerevisiae. The localization of the enzymes involved in the early stage of ergosterol synthesis was investigated. 2. 2. The highest specific activity of 3-hydroxy-3-methylglutaryl-CoA reductase (mevalonate: NADP+ oxidoreductase, EC 1.1.1.34) was associated with the mitochondrial fraction of cells harvested during the late logarithmic and stationary growth phases. The activity of the enzyme in the other fractions was negligible. The distribution ofhydroxymethylglutaryl-CoA reductase was coincident with that of succinate dehydrogenase (EC 1.3.99.1), a mitochondrial marker enzyme. 3. 3. A large part of acetoacetyl-CoA thiolase (acetyl-CoA: acetyl-CoA C-acetyltransferase, EC 2.3.1.9) was present in the cytoplasmic fraction, but a significant part of the enzyme was also present in the mitochondrial fraction. 4. 4. The mitochondrial fraction, in the presence of ATP, Mg2+, CoA, and an NADPH-generating system, could incorporate radioactivity from [1-14C]acetate into mevalonate. 5. 5. No single subcellular fraction incorporated radioactivity from hydroxy [3-14C]methylglutaryl-CoA into nonsaponifiable lipids, whereas a mixture of the mitochondrial and cytoplasmic fractions effected such an incorporation. Only the cytoplasmic fraction incorporated radioactivity from [2-14C]mevalonate into nonsaponifiable lipids.
収録刊行物
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- Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism
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Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism 296 310-320, 1973-02-01
Elsevier BV