Centre for Molecular Cell Biology, UCL, Institute of Liver and Digestive Health, Royal Free Campus, UCL, London NW3 2PF, United Kingdom.
Biol Cell. 2012 Oct;104(10):618-27. doi: 10.1111/boc.201200020. Epub 2012 Jul 24.
In recent years, there has been considerable interest in mapping the protein content of isolated organelles using mass spectrometry. However, many subcellular compartments are highly dynamic with diverse and intricate architectures that are not always preserved during membrane isolation procedures. Furthermore, lateral heterogeneities in intra-membrane lipid and protein concentrations underlie the formation of membrane microdomains, trafficking vesicles and inter-membrane contacts. These complexities in membrane organisation have important consequences for the design of membrane preparation strategies and test the very concept of organelle purity. We illustrate how some of these biological considerations are relevant to membrane preparation and assess the numerous potential pitfalls in attempting to purify organelles from mammalian cells.
近年来，人们对使用质谱法绘制分离细胞器的蛋白质含量图谱产生了浓厚的兴趣。然而，许多亚细胞隔室具有高度动态性，其结构复杂，在膜分离过程中并不总是得到保留。此外，膜内脂质和蛋白质浓度的侧向异质性是膜微区、运输小泡和膜间接触形成的基础。这些膜组织的复杂性对膜制备策略的设计有重要影响，也对细胞器纯度的概念提出了挑战。我们举例说明了其中一些生物学考虑因素与膜制备的相关性，并评估了从哺乳动物细胞中纯化细胞器时可能遇到的众多潜在陷阱。
