1990
DOI: 10.1083/jcb.111.3.941
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Nonselective autophagy of cytosolic enzymes by isolated rat hepatocytes.

Abstract: Abstract. Seven cytosolic enzymes with varying halflives (ornithine decarboxylase, 0.9 h; tyrosine aminotransferase, 3.1 h; tryptophan oxygenase, 3.3 h; serine dehydratase, 10.3 h, glucokinase, 12.7 h; lactate dehydrogenase, 17.0 h; aldolase, 17.4 h) were found to be autophagically sequestered at the same rate (3.5%/h) in isolated rat hepatocytes. Autophagy was measured as the accumulation of enzyme activity in the sedimentable organdies (mostly lysosomes) of electrodisrupted cells in the presence of the prote… Show more

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Cited by 211 publications
(210 citation statements)
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“…6 We simplified the analysis of hepatocellular LC3 dynamics by blocking de novo influx of LC3 with the protein synthesis inhibitor cycloheximide. Under these conditions, total LC3 (LC3-I C LC3-II) declined rapidly (half-life »2 h); 6 this early LC3 disappearance could largely be accounted for by an autophagic-lysosomal degradation that was suppressed by inhibitors of autophagic sequestration (3MA, TG) or lysosomal function (NH 4 Cl, bafilomycin A 1 , leupeptin) 6 (Fig. 1A, inhibitors added at 0 min).…”
Section: Introductionmentioning
confidence: 94%
See 3 more Smart Citations
“…6 We simplified the analysis of hepatocellular LC3 dynamics by blocking de novo influx of LC3 with the protein synthesis inhibitor cycloheximide. Under these conditions, total LC3 (LC3-I C LC3-II) declined rapidly (half-life »2 h); 6 this early LC3 disappearance could largely be accounted for by an autophagic-lysosomal degradation that was suppressed by inhibitors of autophagic sequestration (3MA, TG) or lysosomal function (NH 4 Cl, bafilomycin A 1 , leupeptin) 6 (Fig. 1A, inhibitors added at 0 min).…”
Section: Introductionmentioning
confidence: 94%
“…3 The bulk nature of macroautophagy has been strikingly demonstrated by the fact that cytosolic enzymes with widely different half-lives are all sequestered into autophagic vacuoles at the same rate, which also make them potential probes for macroautophagic cargo sequestration. 4 We routinely use LDH (lactate dehydrogenase) for this purpose, since it is a cytosolic enzyme present in all cells, and is degraded exclusively by the autophagic-lysosomal pathway. 4 Our macroautophagic cargo sequestration assay, which can be adapted to any cell type, 5 measures the net transfer of LDH into sedimentable autophagic vacuoles in the presence of leupeptin or bafilomycin A 1 to prevent LDH degradation in amphisomes and lysosomes.…”
Section: Introductionmentioning
confidence: 99%
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“…It has been assumed for a long time that macroautophagy is a non-selective process, in which macromolecules are randomly degraded in the same ratio as they occur in the cytoplasm (Kominami et al, 1983;Kopitz et al, 1990). However, recent observations strongly suggest that this may not always be the case, and that macroautophagy can be selective under some conditions.…”
Section: Specificity Of Macroautophagymentioning
confidence: 99%