Evaluation of two cell surface modification methods for proteomic analysis of plasma membrane from isolated mouse hepatocytes
Evaluation of two cell surface modification methods for proteomic analysis of plasma membrane from isolated mouse hepatocytes
复制标题
两种细胞表面修饰方法对离体小鼠肝细胞质膜蛋白质组学分析的评价
DOI：
10.1016/j.bbapap.2008.07.009
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发表时间：
2009-01-01
影响因子：
3.2
通讯作者：
Liang, Songping
中科院分区：
文献类型：
--
作者：
Li, Xuanwen;Jin, Qihui;Liang, Songping
肝细胞是一种高度极化的细胞，质膜(PM)蛋白分布不均。为了降低肝组织蛋白质组学的复杂性，获得完整的肝细胞质膜，对生物素-亲和素(BA)和阳离子二氧化硅-聚阴离子(CSP)两种基于细胞表面修饰的PM纯化方法进行了评价，并与传统的细胞分离法进行了比较。采用不同的PM修饰原理，两种方法都能有效地分离纯化的PM组分和普通组分。CSP法从新鲜分离的肝细胞中提纯PM的产率较高。共鉴定出189个非冗余蛋白质，其中49个来自BA方法，185个来自CSP策略。许多已知的和新的PM相关蛋白也被发现。我们的评估应该对从其他新鲜分离的组织来源的细胞制备PM有一定的指导意义。本文所鉴定的肝细胞PM蛋白可作为PM相关功能和疾病研究的参考。(C)2008爱思唯尔B.V.保留所有权利。
The hepatocyte is a highly polarized cell with a heterogeneous distribution of plasma-membrane (PM) proteins. To reduce the complexity of the proteome of liver tissue and give a comprehensive profile of hepatocyte PM, two PM purification methods based on cell Surface modification, named the biotin-avidin (BA) and cationic silica-polyanion (CSP) strategies were evaluated and compared with the traditional cell fractionation method to prepare highly enriched PM from freshly isolated C57 mouse hepatocytes. Employing different principles for PM modification, both methods were effective in the isolation of general and purified PM fraction. The CSP strategy showed better yield for the PM purification from freshly isolated hepatocytes. 189 non-redundant proteins were identified, including 49 from the BA method and 185 from CSP strategy. Many known and novel PM-associated proteins were also found. Our evaluation here should give implications for PM preparation from other freshly isolated tissue-derived cells. The hepatocyte PM proteins identified here Should be taken as a references for the PM-related functional and diseases research. (C) 2008 Elsevier B.V. All rights reserved.