文献类型：期刊文献

英文题名：Enhancing Membrane Protein Identification Using a Simplified Centrifugation and Detergent-Based Membrane Extraction Approach

作者：Zhou, Yanting[1,2,3];Gao, Jing[2,3];Zhu, Hongwen[2,3];Xu, Jingjing[2,3];He, Han[2,3];Gu, Lei[1];Wang, Hui[1];Chen, Jie[2,3];Ma, Danjun[4,5];Zhou, Hu[2,3];Zheng, Jing[1]

机构：[1]East China Univ Sci & Technol, Shanghai Key Lab New Drug Design, Sch Pharm, Shanghai 200237, Peoples R China;[2]Chinese Acad Sci, Shanghai Inst Mat Med, Dept Analyt Chem, Shanghai 201203, Peoples R China;[3]Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China;[4]Dongguan Univ Technol, Coll Mech Engn, Dongguan 523808, Guangdong, Peoples R China;[5]Qingzi Biotechnol Shenzhen LLC, 4026 Shen Nan Middle Rd, Shenzhen 518039, Guangdong, Peoples R China

年份：2018

卷号：90

期号：4

起止页码：2434

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20180804829338);WOS:【SCI-EXPANDED(收录号:WOS:000426143100008)】；

基金：This research was supported by Grant No. 2017YFC1700204 of the National Key Research and Development Program from the Ministry of Science and Technology of China, by Grant No. 21375138 and 31570830 from National Natural Science Foundation of China, by the Strategic Priority Research Program of the Chinese Academy of Science, "Personalized Medicines-Molecular Signature-based Drug Discovery and Development" (No. XDA12030203), by the Innovation Project of Instrument and Equipment Function Development (Grant No. YZ201542, etc.), and the Bureau of Goods, Chinese Academy of Sciences and by Key Laboratory of Receptor Research, Chinese Academy of Sciences. The mass spectrometry proteomics data have been deposited to the ProteomeX-change Consortium via the PRIDE18 partner repository with the dataset identifier PXD008456.

语种：英文

外文关键词：Activation energy - Drug delivery - Activation analysis - Membranes - Extraction - Solubility - Proteins

摘要：Membrane proteins may act as transporters, receptors, enzymes, and adhesion-anchors, accounting for nearly 70% of pharmaceutical drug targets. Difficulties in efficient enrichment, extraction, and solubilization still exist because of their relatively low abundance and poor solubility. A simplified membrane protein extraction approach with advantages of user-friendly sample processing procedures, good repeatability and significant effectiveness was developed in the current research for enhancing enrichment and identification of membrane proteins. This approach combining centrifugation and detergent along with LC-MS/MS successfully identified higher proportion of membrane proteins, integral proteins and transmembrane proteins in membrane fraction (76.6%, 48.1%, and 40.6%) than in total cell lysate (41.6%, 16.4%, and 13.5%), respectively. Moreover, our method tended to capture membrane proteins with high degree of hydrophobicity and number of transmembrane domains as 486 out of 2106 (23.0%) had GRAVY > 0 in membrane fraction, 488 out of 2106 (23.1%) had TMs >= 2. It also provided for improved identification of membrane proteins as more than 60.6% of the commonly identified membrane proteins in two cell samples were better identified in membrane fraction with higher sequence coverage. Data are available via ProteomeXchange with identifier PXD008456.