Membrane Proteome Analysis of Microdissected Ovarian Tumor Tissues Using Capillary Isoelectric Focusing/Reversed-Phase Liquid Chromatography-Tandem MS
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- 作者：Weijie Wang ; Tong Guo ; Paul A. Rudnick ; Tao Song ; Jie Li ; Zhengping Zhuang ; Wenxin Zheng ; Don L. DeVoe ; Cheng S. Lee ; Brian M. Balgley
- 刊名：Analytical Chemistry
- 出版年：2007
- 出版时间：February 1, 2007
- 年：2007
- 卷：79
- 期：3
- 页码：1002 - 1009
- 全文大小：324K
- 年卷期：v.79,no.3(February 1, 2007)
- ISSN：1520-6882
文摘
This work expands our tissue proteome capabilities fromthe analysis of soluble proteins in previous studies to theexamination of membrane proteins within the pellets ofenriched and selectively isolated tumor cells procuredfrom microdissected tissue specimens. The pellets oftargeted ovarian tumor cells are treated by two differentmembrane protein extraction methods, including the useof detergent and organic solvent. The detergent-basedmembrane protein preparation protocol not only extractsproteins effectively from cell pellets but also is compatiblewith subsequent proteome analysis using combined capillary isoelctric focusing/nano reversed-phase liquid chromatography separations coupled with nano electrosprayionization mass spectrometry. Among proteins identifiedfrom an amount of pellet equivalent to 20 000 cells, 773proteins are predicted to contain one or more transmembrane domains, corresponding to 22% membrane proteome coverage within the SwissProt Human proteinsequence entries.