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T细胞中脂筏的分析。

Analysis of lipid rafts in T cells.

作者信息

Thomas Sunil, Preda-Pais Anca, Casares Sofia, Brumeanu Teodor-D

机构信息

Department of Microbiology, Mount Sinai School of Medicine, 1 Gustave L. Levy Place, New York, NY 10029, USA.

出版信息

Mol Immunol. 2004 Jun;41(4):399-409. doi: 10.1016/j.molimm.2004.03.022.

Abstract

The plasma membrane of T cells is made of a combination of glycosphingolipids and protein receptors organized in glycolipoprotein microdomains termed lipid rafts. The structural assembly of lipid rafts was investigated by various physical and biochemical assays. Depending on the differentiation status of T cells, the lipid rafts seclude various protein receptors involved in T cell signaling, cytoskeleton reorganization, membrane trafficking, and the entry of infectious organisms into the cells. This review article summarizes the most common methods, and their limits and advantages for analyzing the composition and assembly of lipid rafts with protein receptors into lipid rafts microdomains in plasma membrane of T cells. It also includes new methods such as ELISA/Polysorp and flow cytometry, and a combined sucrose gradient centrifugation-FPLC-Western blot strategy developed in our laboratory to study non-covalent interactions between the GM1 glycosphingolipid and protein receptors in plasma membrane of T cells.

摘要

T细胞的质膜由糖鞘脂和蛋白质受体组合而成,这些成分组织在称为脂筏的糖脂蛋白微结构域中。通过各种物理和生化分析方法对脂筏的结构组装进行了研究。根据T细胞的分化状态,脂筏会隔离参与T细胞信号传导、细胞骨架重组、膜运输以及感染性生物体进入细胞过程的各种蛋白质受体。这篇综述文章总结了分析T细胞质膜中脂筏与蛋白质受体组成脂筏微结构域的最常用方法,以及这些方法的局限性和优点。它还包括酶联免疫吸附测定/多吸附酶联免疫吸附测定(ELISA/Polysorp)和流式细胞术等新方法,以及我们实验室开发的一种蔗糖密度梯度离心-FPLC-蛋白质免疫印迹联合策略,用于研究T细胞质膜中GM1糖鞘脂与蛋白质受体之间的非共价相互作用。

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