Microsomal fractions, both homogeneous in appearance and functionally operative, were isolated from a homogenate of rat cerebral cortex by fractionation in water. The preparations thus obtained contain the membranous elements of the endoplasmic reticulum, synaptic vesicles, and ribosomes. Esterase, ATPase, and glutamine synthetase were found to be present and fully functional in the microsomal fractions isolated in water. The contamination of the water-isolated microsomal fractions by mitochondria and lysosomes was found to be considerably lower than in microsomal pellets isolated in sucrose. The contamination by nerve ending particles, as judged by electron microscopy and by the levels of soluble lactic dehydrogenase entrapped in the cytoplasm of the particles, was also low. Most of the contamination by mitochondria and nerve ending particles could be removed by treatment of the microsomal pellet with 150 mM NaCl. Resistant to elution by this treatment is the lysosomal contamination as well as microsomal esterase and ATPase. Glutamine synthetase, on the other hand, was almost totally solubilized. Microsomal preparations isolated in water are also shown to contain amounts of protein, RNA, phospholipid, and ganglioside comparable to those found in microsomal preparations isolated in sucrose.
MORPHOLOGICAL AND BIOCHEMICAL CORRELATES OF CEREBRAL MICROSOMES : I. Isolation and Chemical Characterization
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F. de Balbian Verster, O. Z. Sellinger, J. C. Harkin; MORPHOLOGICAL AND BIOCHEMICAL CORRELATES OF CEREBRAL MICROSOMES : I. Isolation and Chemical Characterization . J Cell Biol 1 May 1965; 25 (2): 69–80. doi: https://doi.org/10.1083/jcb.25.2.69
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