Abstract
Low-density lipoproteins (LDLs) are taken up by the cell mainly through receptor-mediated endocytosis. LDL-derived cholesterol leaves lysosome and further transports to downstream organelles for specific cellular needs. We recently report that cholesterol transfers from lysosome to peroxisome through lysosome–peroxisome membrane contact (LPMC). Here, we use iodixanol density gradient centrifugation to isolate lysosomes and peroxisomes separately for the in vitro reconstitution of LPMC. We also apply 3H-cholesterol-labeled lysosomes and peroxisomes in vitro to measure 3H-cholesterol transfer through LPMC.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (31430044, 31230020, and 81270155 to B.-L.S. and 31600651 to J.L.) and the Natural Science Foundation of Hubei Province (2015CFB215 to J.L.).
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Luo, J., Liao, YC., Xiao, J., Song, BL. (2017). Measurement of Cholesterol Transfer from Lysosome to Peroxisome Using an In Vitro Reconstitution Assay. In: Gelissen, I., Brown, A. (eds) Cholesterol Homeostasis. Methods in Molecular Biology, vol 1583. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6875-6_11
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DOI: https://doi.org/10.1007/978-1-4939-6875-6_11
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