A novel approach to examining compositional heterogeneity of detergent-resistant lipid rafts
A novel approach to examining compositional heterogeneity of detergent-resistant lipid rafts
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DOI：
10.1111/j.1440-1711.2006.01421.x
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发表时间：
2006-04-01
影响因子：
4
通讯作者：
Bamezai, A
中科院分区：
文献类型：
--
作者：
George, S;Nelson, MD;Bamezai, A
脂筏在细胞信号传导、细胞粘附和其他细胞功能中起重要作用。脂筏的组成异质性提供了脂筏如何提供细胞信号传导和细胞粘附的空间和时间调节的一种机制。一些信号传导受体/分子的组成性存在和其他信号传导受体/分子在脂筏中的积累允许它们彼此相互作用，从而促进信号从质膜传递到细胞内部。设计一种方法，可以分析这些脂质微结构域的存在下，信号受体/分子的一个单独的筏的基础上，需要解决的问题，脂筏异质性。SDS-PAGE分析，目前用于分析耐洗涤剂的脂筏，没有解决这个问题。我们设计了一种无细胞检测方法，该方法用抗脂筏驻留分子的抗体捕获耐洗涤剂的脂筏，并检测另一种脂筏分子的存在。我们的研究结果表明，抗洗涤剂的脂筏，也被称为抗洗涤剂膜，是异质群体的永生化小鼠T细胞质膜上的抗原受体/信号复合物和其他信号/粘附蛋白。这种无细胞的测定提供了一种简单而快速的方法来检查同时存在的两种蛋白质的脂筏，并有可能估计贩运的分子进出的脂质微结构域在细胞信号传导的一个单一的洗涤剂耐脂筏的基础上。
Lipid rafts play an important role in cell signalling, cell adhesion and other cellular functions. Compositional heterogeneity of lipid rafts provides one mechanism of how lipid rafts provide the spatial and temporal regulation of cell signalling and cell adhesion. The constitutive presence of some signalling receptors/molecules and accumulation of others in the lipid raft allows them to interact with each other and thereby facilitate relay of signals from the plasma membrane to the cell interior. Devising a method that can analyze these lipid microdomains for the presence of signalling receptors/molecules on an individual raft basis is required to address the issue of lipid raft heterogeneity. SDS-PAGE analysis, currently used for analyses of detergent-resistant lipid rafts, does not address this question. We have designed a cell-free assay that captures detergent-resistant lipid rafts with an antibody against a raft-resident molecule and detects the presence of another lipid raft molecule. Our results suggest that detergent-resistant lipid rafts, also known as detergent-resistant membranes, are heterogeneous populations on an immortalized mouse T-cell plasma membrane with respect to antigen receptor/signalling complex and other signalling/adhesion proteins. This cell-free assay provides a simple and quick way to examine the simultaneous presence of two proteins in the lipid rafts and has the potential to estimate trafficking of molecules in and out of the lipid microdomains during cell signalling on a single detergent-resistant lipid raft basis.