期刊
NATURE PROTOCOLS
卷 9, 期 2, 页码 312-322出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2014.016
关键词
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资金
- Polish National Science Centre [UMO-2011/01/M/NZ3/02128]
- Iuventus Plus [UMO-0531/IP1/2011/71]
- Polish Ministry of Science and Higher Education [W100/HFSC/2011]
- Italian Association for Cancer Research
- Telethon [GGP11139B]
- Italian Ministry of Education, University and Research
- Italian Ministry of Health
- Foundation for Polish Science (FNP)
- EU
- European Regional Development Fund
- 'Innovative economy' Operational Programme
- Polish Ministry of Science and Higher Education
Dynamic interplay between intracellular organelles requires a particular functional apposition of membrane structures. The organelles involved come into close contact, but do not fuse, thereby giving rise to notable microdomains; these microdomains allow rapid communication between the organelles. Plasma membrane-associated membranes (PAMs), which are microdomains of the plasma membrane (PM) interacting with the endoplasmic reticulum (ER) and mitochondria, are dynamic structures that mediate transport of proteins, lipids, ions and metabolites. These structures have gained much interest lately owing to their roles in many crucial cellular processes. Here we provide an optimized protocol for the isolation of PAM, PM and ER fractions from rat liver that is based on a series of differential centrifugations, followed by the fractionation of crude PM on a discontinuous sucrose gradient. The procedure requires similar to 8-10 h, and it can be easily modified and adapted to other tissues and cell types.
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