Plasma membranes from candida tropicalis grown on glucose or hexadecane. I. Isolation,identification and purification |
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Authors: | Heinz Schneider Armin Fiechter Günter F. Fuhrmann |
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Affiliation: | 1. Institute of Microbiology, Swiss Federal Institute of Technology, CH-8092 Zürich Switzerland;2. Department of Pharmacology and Toxicology, School of Medicine, Lahnberge, D-3550 Marburg G.F.R. |
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Abstract: | Plasma membranes from Candida tropicalis grown on glucose or hexadecane were isolated using a method based on the difference in surface charge of mitochondria and plasma membranes.After mechanical disruption of the cells, a fraction consisting of mitochondrial and plasma membrane vesicles was obtained by differential centrifugation.Subsequently the mitochondria were separated from the plasma membrane vesicles by aggregation of the mitochondria at a pH corresponding to their isoelectric point. Additional purification of the isolated plasma membrane vesicles was achieved by osmolysis. Surface charge densities of mitochondria and plasma membranes were determined and showed substrate-dependent differences.The isolated plasma membranes were morphologically characterized by electron microscopy and, as a marker enzyme, the activity of Mg2+-dependant ATPase was determined.By checking for three mitochondrial marker enzymes the plasma membrane fractions were estimated to be 94% pure with regard to mitochondrial contamination. |
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