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- W243559497 abstract "Chaperone-mediated autophagy (CMA) is a lysosomal pathway of proteolysis that degrades 30% of cytosolic proteins during nutritional deprivation. Substrates for this pathway of proteolysis have targeting sequences biochemically related to the pentapeptide KFERQ. Another lysosomal pathway of proteolysis, macroautophagy, is induced in rat liver for the first day of starvation, but macroautophagy declines to basal levels by the second day. Once the cells are ready to harvest, usually at confluence, media can be removed from the plate using a vacuum flask and Pasteur pipette. Preparation of lysosomes involves resuspending the pelleted cells in 1 ml of sucrose solution and placing them into a nitrogen cavitation chamber. The latency assay is necessary to determine the percentage of broken lysosomes after isolation. Throughout the entire lysosomal preparation the samples should be kept on ice. Failure to keep the samples on ice will significantly decrease lysosomal recovery and increase the percentage of lysosomes that are broken. An 80% Mtz stock solution (w/v) should be made several days prior to the lysosomal preparation." @default.
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- W243559497 date "2006-01-01" @default.
- W243559497 modified "2023-10-16" @default.
- W243559497 title "Protein Degradation MethodsChaperone-Mediated Autophagy" @default.
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- W243559497 doi "https://doi.org/10.1016/b978-012164730-8/50229-x" @default.
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