A simple procedure for the isolation of rat kidney lysosomes.
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.


CONCLUSIONS
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.
Kawashima
A
A
Department of Medicine, Tokyo Women's Medical College, Japan.
Sato
A
A
Kawashima
M
M
Nitta
K
K
Yumura
W
W
Sugino
N
N
Nihei
H
H
Natori
Y
Y
D000418:Albumins / Q000032:analysis; D000818:Animals; D001192:Arylsulfatases / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D002402:Cathepsin D / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods*; D002498:Centrifugation; D003102:Colloids; D042963:Electron Transport Complex II; D005952:Glucose-6-Phosphatase / Q000378:metabolism; D007668:Kidney / Q000166:cytology*; D008247:Lysosomes / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D009097:Multienzyme Complexes / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D011205:Povidone; D051381:Rats; D017208:Rats, Wistar; D012822:Silicon Dioxide; D013385:Succinate Dehydrogenase / Q000378:metabolism
D000418:Albumins; D003102:Colloids; D009097:Multienzyme Complexes; C016039:Percoll; D012822:Silicon Dioxide; D010088:Oxidoreductases; D002374:Catalase; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase; D005952:Glucose-6-Phosphatase; D001192:Arylsulfatases; D002402:Cathepsin D; D011205:Povidone