N-acetyl-beta-D-glucosaminidase (NAG) isoenzymes in primary cultures of rabbit kidney proximal tubule cells: a cellular model for studies on nephrotoxicity?
N-Acetyl-beta-D-glucosaminidase (NAG) isoenzyme profile in primary cultures of rabbit kidney proximal tubule cells was studied. Confluent cells had high levels of NAG activity, but ion exchange chromatography showed that the NAG isoenzyme profile in cultured cells was different from that of rabbit renal cortex homogenates and freshly isolated cells. Confluent cultured cells contained an atypical acidic isoform, absent in homogenates and freshly isolated cells in which the predominant isoform is NAG-A (a heterodimer alpha beta). The fact that this atypical isoform was able to hydrolyse the synthetic substrate 4-methylumbelliferyl-beta-N-acetylglucosaminide-6-sulphate indicated that it probably was an alpha-subunit homodimer. These results suggest subunit rearrangement within NAG polypeptide chains linked to down-regulation of beta-subunit production in cultured rabbit proximal cells. The change in isoenzyme profile in cultured cells may make it difficult to use primary cultures of rabbit proximal tubule cells to establish correlations between in vitro and in vivo studies using NAG isoenzymes as a nephrotoxicity index, as illustrated by the effects of gentamicin.
Laboratory of Biochemistry and Glycobiology, Faculty of Pharmaceutical and Biological Sciences, Université René Descartes, Paris, France.
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D
Robic
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Bens
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Loko
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Vandewalle
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Bourbouze
D000118:Acetylglucosaminidase / Q000378:metabolism*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000008:administration & dosage / Q000633:toxicity; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D015536:Down-Regulation; D005839:Gentamicins / Q000008:administration & dosage / Q000633:toxicity; D006868:Hydrolysis; D007527:Isoenzymes / Q000378:metabolism*; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D011500:Protein Synthesis Inhibitors / Q000008:administration & dosage / Q000633:toxicity; D011817:Rabbits
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D007527:Isoenzymes; D011500:Protein Synthesis Inhibitors; D000118:Acetylglucosaminidase