Rapid separation of particulate components and soluble cytoplasm of isolated rat-liver cells.
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.
Zuurendonk
P F
PF
Laboratory of Biochemistry, B.C.P. Jansen Institute, University of Amsterdam, Plantage Muidergracht 12, Amsterdam, The Netherlands.
Tager
J M
JM
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D003593:Cytoplasm / Q000737:chemistry*; D004072:Digitonin / Q000737:chemistry; D022781:Hepatocytes / Q000737:chemistry* / Q000166:cytology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008930:Mitochondria, Liver / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism
D007770:L-Lactate Dehydrogenase; D004072:Digitonin