title: Buffer-induced rat liver mitochondrial aggregation during differential centrifugation.
abstract: Use of buffers in homogenization media can result in loss of considerable particulate enzyme activity even with low-speed centrifugation. Addition of tris chloride buffer to 0.25 M sucrose homogenization media resulted in precipitation of 80 to 95% of the activity of two mitochondrial marker enzymes (3-hydroxy-3-methylglutaryl CoA lyase and citrate synthase) with the nuclear fraction during differential centrifugation. Lactate dehydrogenase, a cytoplasmic marker, was not precipitated under the same conditions, indicating that the precipitated enzymes were not associated with intact cells. Photomicrographs showed that tris chloride buffers resulted in mitochondrial aggregation. Isolated mitochondria resuspended in tris chloride or potassium phosphate buffer also aggregated, which resulted in a marked decrease in assayable mitochondrial enzyme activity.
mesh_terms: D000818:Animals; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000201:enzymology; D002950:Citrate (si)-Synthase / Q000032:analysis; D003600:Cytosol / Q000201:enzymology; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000032:analysis; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008099:Liver / Q000201:enzymology; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology / Q000648:ultrastructure*; D051381:Rats; D011919:Rats, Inbred Strains; D014461:Ultracentrifugation
chemical_list: D007770:L-Lactate Dehydrogenase; D002950:Citrate (si)-Synthase; D006904:Hydroxymethylglutaryl-CoA Synthase