Epidermal growth factor receptors are localized to lipid rafts that contain a balance of inner and outer leaflet lipids - A shotgun lipidomics study
Epidermal growth factor receptors are localized to lipid rafts that contain a balance of inner and outer leaflet lipids - A shotgun lipidomics study
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DOI：
10.1074/jbc.m503805200
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发表时间：
2005-07-22
影响因子：
4.8
通讯作者：
Gross, RW
中科院分区：
文献类型：
--
作者：
Pike, LJ;Han, XL;Gross, RW
表皮生长因子（EGF）受体分区到脂筏使用无去污剂的方法，但从低密度馏分中提取的Triton X-100。通过筛选几种洗涤剂，我们确定Brij 98作为洗涤剂，其中EGF受体保留在洗涤剂抗性膜组分中。为了确定脂质组成之间的差异筏窝藏EGF受体（清洁剂和Brij 98耐）和那些没有（Triton X-100耐），我们使用多维电喷雾电离质谱进行脂质组学研究这三个筏制剂。虽然所有三个筏制剂类似地富含胆固醇，含有EGF受体的筏含有更多的乙醇胺甘油磷脂和更少的鞘磷脂比不含EGF受体的Triton X-100筏。结果，无洗涤剂和Brij 98抗性筏表现出内外小叶脂质的平衡，而Triton X-100筏含有外小叶脂质的优势。此外，在所有筏准备，外叶磷脂物质显着不同的散装膜，而内叶脂质是非常相似的散装膜中发现的。这些研究结果表明，EGF受体被保留在筏，表现出与双层结构相容的脂质分布和选择磷脂纳入筏主要发生在外部小叶脂质。
The epidermal growth factor (EGF) receptor partitions into lipid rafts made using a detergent-free method, but is extracted from low density fractions by Triton X-100. By screening several detergents, we identified Brij 98 as a detergent in which the EGF receptor is retained in detergent-resistant membrane fractions. To identify the difference in lipid composition between those rafts that harbored the EGF receptor (detergent-free and Brij 98-resistant) and those that did not (Triton X-100-resistant), we used multidimensional electrospray ionization mass spectrometry to perform a lipidomics study on these three raft preparations. Although all three raft preparations were similarly enriched in cholesterol, the EGF receptor-containing rafts contained more ethanolamine glycerophospholipids and less sphingomyelin than did the non-EGF receptor-containing Triton X-100 rafts. As a result, the detergent-free and Brij 98-resistant rafts exhibited a balance of inner and outer leaflet lipids, whereas the Triton X-100 rafts contained a preponderance of outer leaflet lipids. Furthermore, in all raft preparations, the outer leaflet phospholipid species were significantly different from those in the bulk membrane, whereas the inner leaflet lipids were quite similar to those found in the bulk membrane. These findings indicate that the EGF receptor is retained only in rafts that exhibit a lipid distribution compatible with a bilayer structure and that the selection of phospholipids for inclusion into rafts occurs mainly on the outer leaflet lipids.