McClellan Mark E, Elliott Michael H
Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA.
Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, 73104, OK, USA.
Methods Mol Biol. 2017;1609:185-194. doi: 10.1007/978-1-4939-6996-8_16.
The compartmentalization of cellular membranes into discrete membrane microdomains (known as lipid rafts) challenged the original definition of membranes as containing randomly distributed lipid and protein components. The lipid microdomain hypothesis has generated significant controversy and rigorous inquiry based on the attractive idea that such domains concentrate machinery to mediate cellular events such as signaling and endocytosis. As such, numerous studies have used biochemical, cell biological, and biophysical methodologies to define the composition of such domains in a variety of experimental contexts. In this chapter, we describe methodologies to isolate membranes from cell or tissue sources with biophysical/biochemical properties of membrane microdomains that are amenable to subsequent classical or mass spectrometry-based lipid analytical approaches.
细胞膜分隔成离散的膜微区（即脂筏），这对膜的原始定义提出了挑战，原始定义认为膜包含随机分布的脂质和蛋白质成分。脂质微区假说基于这样一个引人注目的观点引发了重大争议和严格探究，即这些微区集中了介导细胞事件（如信号传导和内吞作用）的机制。因此，许多研究已使用生物化学、细胞生物学和生物物理方法来在各种实验环境中定义这些微区的组成。在本章中，我们描述了从细胞或组织来源分离具有膜微区生物物理/生物化学特性的膜的方法，这些特性适合后续基于经典或质谱的脂质分析方法。
