Rapid separation of particulate components and soluble cytoplasm of isolated rat-liver cells |
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Authors: | P.F. Zuurendonk J.M. Tager |
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Affiliation: | Laboratory of Biochemistry, B. C. P. Jansen Institute, University of Amsterdam, Plantage Muidergracht 12, Amsterdam The Netherlands |
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Abstract: | A method is described for the rapid separation of mitochondria (plus other particulate components) from the soluble cytoplasm of isolated rat-liver cells. The cells were incubated briefly with a low concentration of digitonin. After rapid centrifugation, the pellet contained more than 90% of the total adenylate kinase and glutamate dehydrogenase activities and the supernatant at least 80% of the lactate dehydrogenase activity. About 60% of total adenine nucleotides in hepatocytes were found in the soluble cytoplasm. The ATP/ADP ratio in the particulate fraction 80 s after exposure to digitonin of hepatocytes metabolizing alanine was 2.0–2.4, and that in the soluble cytoplasm 6–19. In the presence of atractyloside, these values were 3.5–4.4 and 1.3–2.2, respectively. |
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Keywords: | MOPS morpholinopropane sulphonic acid |
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