Arabidopsis plasma membrane proteomics identifies components of transport, signal transduction and membrane trafficking
Arabidopsis plasma membrane proteomics identifies components of transport, signal transduction and membrane trafficking
复制标题
DOI：
10.1093/pcp/pch209
复制
发表时间：
2004-11-01
影响因子：
4.9
通讯作者：
Kjellbom, P
中科院分区：
文献类型：
--
作者：
Alexandersson, E;Saalbach, G;Kjellbom, P
为了鉴定与质膜相关的整合蛋白和外周蛋白，通过质谱分析来自绿色组织（叶和叶柄）的高度纯化的拟南芥质膜。质膜分离的水两相分配，产生质膜囊泡与细胞质侧的方向和纯度为95%。通过用Brij 58处理将这些囊泡从内到外翻转，以除去囊泡中包封的可溶性污染蛋白质并除去松散结合的污染蛋白质。总共鉴定了238种推定的质膜蛋白，其中114种被预测具有跨膜结构域或被糖基磷脂酰肌醇锚定。约三分之二的已鉴定的整合蛋白以前没有被证明是质膜蛋白。在238个鉴定的蛋白质中，76%可以根据功能进行分类。主要类别是参与运输（17%），信号转导（16%），膜运输（9%）和应激反应（9%）的蛋白质。在本研究中鉴定的蛋白质中，几乎有四分之一在功能上未分类，其中一半以上预计是不可分割的。
In order to identify integral proteins and peripheral proteins associated with the plasma membrane, highly purified Arabidopsis plasma membranes from green tissue (leaves and petioles) were analyzed by mass spectrometry. Plasma membranes were isolated by aqueous two-phase partitioning, which yields plasma membrane vesicles with a cytoplasmic-side-in orientation and with a purity of 95%. These vesicles were turned inside-out by treatment with Brij 58 to remove soluble contaminating proteins enclosed in the vesicles and to remove loosely bound contaminating proteins. In total, 238 putative plasma membrane proteins were identified, of which 114 are predicted to have transmembrane domains or to be glycosyl phosphatidylinositol anchored. About two-thirds of the identified integral proteins have not previously been shown to be plasma membrane proteins. Of the 238 identified proteins, 76% could be classified according to function. Major classes are proteins involved in transport (17%), signal transduction (16%), membrane trafficking (9%) and stress responses (9%). Almost a quarter of the proteins identified in the present study are functionally unclassified and more than half of these are predicted to be integral.