Measurement of Cholesterol Transfer from Lysosome to Peroxisome Using an In Vitro Reconstitution Assay

Jie Luo; Ya-Cheng Liao; Jian Xiao; Bao-Liang Song

doi:10.1007/978-1-4939-6875-6_11

Measurement of Cholesterol Transfer from Lysosome to Peroxisome Using an In Vitro Reconstitution Assay

Methods Mol Biol. 2017:1583:141-161. doi: 10.1007/978-1-4939-6875-6_11.

Authors

Jie Luo¹, Ya-Cheng Liao², Jian Xiao¹, Bao-Liang Song³

Affiliations

¹ Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, 430072, China.
² State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
³ Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan, 430072, China. [email protected].

PMID: 28205172
DOI: 10.1007/978-1-4939-6875-6_11

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.

Keywords: Cholesterol transfer; In vitro reconstitution assay; Lysosome; Lysosome–peroxisome membrane contact; Peroxisome.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biological Transport, Active / physiology
Cholesterol* / chemistry
Cholesterol* / metabolism
HeLa Cells
Humans
Intracellular Membranes* / chemistry
Intracellular Membranes* / metabolism
Lysosomes* / chemistry
Lysosomes* / metabolism
Peroxisomes* / chemistry
Peroxisomes* / metabolism

Substances

Cholesterol