The isolation and structure of membrane lipid rafts from rat brain
The isolation and structure of membrane lipid rafts from rat brain
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DOI：
10.1016/j.biochi.2006.07.006
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发表时间：
2007-02-01
期刊：
影响因子：
3.9
通讯作者：
Quinn, P. J.
中科院分区：
文献类型：
--
作者：
Chen, X.;Morris, R.;Quinn, P. J.
本文报道了洗涤剂在分离大鼠脑抗洗涤剂膜组分中的作用。Triton X-100在4摄氏度下处理全大鼠脑匀浆，产生了密度为1.07 g/ml的耐洗涤剂膜，而Brij 96的膜密度仅为1.05 g/ml。使用Triton X-100分离的DRM级分比从用Brij 96处理的匀浆中分离的那些重得多。与Triton X-100衍生的DRM相比，Brij 96处理的匀浆衍生的DRM的主要极性脂质组成具有比胆碱磷脂更高比例的氨基磷脂;这可能表明Brij 96处理的匀浆衍生的DRM与脂质组成中的母膜更密切相关。Triton X-100在较高温度下的溶解导致第二耐洗涤剂膜部分的出现，其密度明显轻于在密度1.07 g/ml下回收的膜。在37 ℃下洗涤剂处理长达30分钟期间，脑匀浆的磷脂组成分析显示鞘磷脂的比例降低。在37 ℃下处理匀浆似乎激活磷脂酶/鞘磷脂酶，这可能会改变分离的DRM的脂质含量。K+/Mg 2+与Brij 96处理的存在下，结果在DRM馏分具有显着更厚的双层和更大的囊泡直径比DRM分离的Triton X-100或Brij 96处理的匀浆在阳离子的情况下。(c)2006年，Elsevier Masson SAS。All rights reserved.
The action of detergents in the isolation of detergent-resistant membrane fractions from rat brain is reported. Triton X-100 treatment of whole rat brain homogenate at 4 degrees C produced detergent-resistant membranes with a density of 1.07 g/ml compared with Brij96 where the density of the membrane was only 1.05 g/ml. The DRM fractions isolated using Triton X-100 are considerably heavier than those isolated from homogenates treated with Brij96. The major polar lipid composition of DRMs derived from Brij96 treated homogenates have a higher proportion of amino-phospholipids compared with choline phospholipids than Triton X-100 derived DRMs; this may indicate that DRMs from Brij96 treated homogenates are more closely related to the parent membrane in lipid composition. Solubilization by Triton X-100 at higher temperatures resulted in the appearance of a second detergent-resistant membrane fraction distinctly lighter in density than the membrane recovered at density 1.07 g/ml. Analysis of phospholipid composition of the brain homogenate during detergent treatment for up to 30 min at 37 degrees C showed a decreasing proportion of sphingomyelin. Treatment of homogenates at 37 degrees C appears to activate phospholipases/sphingomyelinases that may alter the lipid content of isolated DRMs. The presence of K+/Mg2+ with Brij96 treatment results in DRM fractions with significantly thicker bilayers and of larger vesicle diameter than DRMs isolated from either Triton X-100 or Brij96 treated homogenates in the absence of cations. (c) 2006 Elsevier Masson SAS. All rights reserved.