A simple procedure for the isolation of rat kidney lysosomes

Kidney Int. 1998 Jul;54(1):275-8. doi: 10.1046/j.1523-1755.1998.00958.x.

Abstract

Background: A procedure for the isolation of highly purified lysosomes from normal rat kidney is described.

Methods: The method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 2 mM Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation.

Results: The lysosomal fraction obtained by our method was enriched more than 30-fold in terms of marker enzymes with a yield of about 11%. Electron microscopic examination and the measurement of the activities of marker enzymes for various subcellular organelles indicated that our lysosomal preparation was essentially free from contamination by other organelles.

Conclusion: We believe that this procedure for isolating kidney lysosome will be useful in the study of the mechanisms of specific modification, processing and catabolism of proteins.

Publication types

  • Technical Report

MeSH terms

  • Albumins / analysis
  • Animals
  • Arylsulfatases / metabolism
  • Catalase / metabolism
  • Cathepsin D / metabolism
  • Cell Fractionation / methods*
  • Centrifugation
  • Colloids
  • Electron Transport Complex II
  • Glucose-6-Phosphatase / metabolism
  • Kidney / cytology*
  • Lysosomes / chemistry
  • Lysosomes / enzymology
  • Lysosomes / ultrastructure*
  • Male
  • Microscopy, Electron
  • Multienzyme Complexes / metabolism
  • Oxidoreductases / metabolism
  • Povidone
  • Rats
  • Rats, Wistar
  • Silicon Dioxide
  • Succinate Dehydrogenase / metabolism

Substances

  • Albumins
  • Colloids
  • Multienzyme Complexes
  • Percoll
  • Silicon Dioxide
  • Oxidoreductases
  • Catalase
  • Electron Transport Complex II
  • Succinate Dehydrogenase
  • Glucose-6-Phosphatase
  • Arylsulfatases
  • Cathepsin D
  • Povidone