title: Measurement of Cholesterol Transfer from Lysosome to Peroxisome Using an In Vitro Reconstitution Assay.
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 apply3H-cholesterol-labeled lysosomes and peroxisomes in vitro to measure3H-cholesterol transfer through LPMC.
authors.affiliation: Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, 430072, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
authors.forename: Ya-Cheng
authors.initials: YC
authors.lastname: Liao
authors.affiliation: Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, 430072, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Xiao
authors.affiliation: Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, 430072, China. blsong@whu.edu.cn.
authors.forename: Bao-Liang
authors.initials: BL
authors.lastname: Song
mesh_terms: D001693:Biological Transport, Active / Q000502:physiology; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism; D008247:Lysosomes / Q000737:chemistry / Q000378:metabolism; D020675:Peroxisomes / Q000737:chemistry / Q000378:metabolism
chemical_list: D002784:Cholesterol
keywords: Cholesterol transfer; In vitro reconstitution assay; Lysosome; Lysosome–peroxisome membrane contact; Peroxisome