Document Detail

Analytical cell fractionation of isolated rabbit renal proximal tubules.
MedLine Citation:
PMID:  7300116     Owner:  NLM     Status:  MEDLINE    
Proximal tubules were isolated in highly pure form from rabbit cortices by a mechanical procedure that is known to preserve the structural and metabolic aspects of the tubular cells. Postnuclear supernates prepared from the isolated tubules were subjects to isopycnic centrifugation in linear sucrose gradients. The enzyme activities associated with the plasma membrane (gamma-glutamyl transpeptidase, amino-peptidase M, alkaline phosphatase, Na-K-ATPase, and phosphodiesterase I) exhibited sharp unimodal frequency-density profiles with a median density near 1.16 g/ml, which shifted to a heavier density when treated with digitonin. The lysosomal enzymes, N-acetyl-beta-glucosaminidase, alpha-mannosidase, and cathepsin B, and the peroxisomal enzyme catalase exhibited particle-associated activity near a density of 1.22 g/ml. Disruption of these particles by freezing and thawing resulted in these activities appearing in the rho = 1.10 g/ml region of the gradient where the soluble cytosolic enzyme, phosphoglucomutase, exhibited activity. Cytochrome oxidase activity typical of mitochondria gave a sharp unimodal profile at rho = 1.18 g/ml. Microsomal glucose-6-phosphatase and NADPH: cytochrome c reductase activities gave median densities near 1.16 g/ml, which did not change after incubation with digitonin. Galactosyl transferase activity gave a skewed profile at rho = 1.16 g/ml and showed a slight shift to heavier density after digitonin. This study of the enzymatic activities and density gradient distribution of the components of the proximal tubule cells provides the methodology for the further study of the cellular processing of endogenous and exogenous substances by this vital cell type.
J T Hjelle; J P Morin; A Trouet
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Kidney international     Volume:  20     ISSN:  0085-2538     ISO Abbreviation:  Kidney Int.     Publication Date:  1981 Jul 
Date Detail:
Created Date:  1982-01-09     Completed Date:  1982-01-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0323470     Medline TA:  Kidney Int     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  71-7     Citation Subset:  IM    
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MeSH Terms
Cell Fractionation
Cell Membrane / enzymology
Kidney Tubules, Proximal / cytology*
Lysosomes / enzymology
Organoids / enzymology
Subcellular Fractions / enzymology

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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