SWATH-Based Comprehensive Determination of the Localization of Apical and Basolateral Membrane Proteins Using Mouse Liver as a Model Tissue.
SWATH-Based Comprehensive Determination of the Localization of Apical and Basolateral Membrane Proteins Using Mouse Liver as a Model Tissue.
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DOI：
10.3390/biomedicines10020383
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发表时间：
2022-02-05
期刊：
影响因子：
4.7
通讯作者：
Uchida Y
中科院分区：
文献类型：
--
作者：
Hirano S;Goto R;Uchida Y
本研究的目的是开发一种方法来全面确定顶膜和基底外侧膜蛋白的定位，使用顶/基底外侧膜分离和准确的SWATH（全理论碎片离子谱的顺序窗口采集）蛋白质组学的组合。SWATH分析小鼠肝脏中基底外侧和顶端质膜组分定量了1373种蛋白质的蛋白质表达。蛋白质表达水平的基底/顶端比率与41种模型蛋白质（23种基底外侧蛋白质，11种顶端蛋白质和7种膜定位蛋白质）的免疫组织化学定位进行了比较。三个群体完全不同。基于概率密度，确定区分顶侧、两侧和基底侧定位的边界线为0.766和1.42。将建立的方法应用于小鼠肝脏中蛋白质的综合测定。结果表明，154和125蛋白定位在顶膜和基底侧膜，分别。顶膜中受体、CD抗原和整合素、酶和Ras相关分子的水平远高于基底侧膜。相反，基底外侧膜中的粘附分子、支架蛋白和转运蛋白的水平远高于顶膜。
The purpose of this study was to develop a method to comprehensively determine the localization of apical and basolateral membrane proteins, using a combination of apical/basolateral membrane separation and accurate SWATH (Sequential Window Acquisition of all THeoretical fragment ion spectra) proteomics. The SWATH analysis of basolateral and apical plasma membrane fractions in mouse liver quantified the protein expression of 1373 proteins. The basolateral/apical ratios of the protein expression levels were compared with the reported immunohistochemical localization for 41 model proteins (23 basolateral, 11 apical and 7 both membrane-localized proteins). Three groups were perfectly distinguished. Border lines to distinguish the apical-, both- and basolateral localizations were determined to be 0.766 and 1.42 based on probability density. The method that was established was then applied to the comprehensive determination of the proteins in mouse liver. The findings indicated that 154 and 125 proteins were localized in the apical and basolateral membranes, respectively. The levels of receptors, CD antigens and integrins, enzymes and Ras-related molecules were much higher in apical membranes than in basolateral membranes. In contrast, the levels of adhesion molecules, scaffold proteins and transporters in basolateral membranes were much higher than in apical membranes.
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DOI：
10.1152/ajpgi.00492.2005
发表时间：
2006-05-01
影响因子：
4.5
作者：
Quondamatteo, F;Krick, W;Burckhardt, BC
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影响因子：
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影响因子：
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DOI：
10.1073/pnas.0904411106
发表时间：
2009-09-08
影响因子：
11.1
作者：
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通讯作者：
Thorens, Bernard