Evaluation of strategy for analyzing mouse liver plasma membrane proteome

• Published in: Science China. Life sciences 2007-12, Vol.50 (6), p.731-738
• Main Authors: Chen, Ping, Zhang, LiJun, Li, XuanWen, Wang, XiE, Cao, Rui, Liu, Zhen, Xiong, JiXian, Peng, Xia, Wei, YingJuan, Ying, XingFeng, Wang, XianChun, Liang, SongPing
• Format: Article
• Language: English
• Subjects: Animals; Cell Fractionation; Cell Membrane - chemistry; Centrifugation; Contamination; Liver; Liver - chemistry; Liver Extracts - analysis; Liver Extracts - chemistry; Mass spectroscopy; Membrane Proteins - analysis; Membrane Proteins - isolation & purification; Mice; Mice, Inbred C57BL; Mitochondria; Proteome - analysis; Proteome - chemistry; Proteomes; Trypsin; Western blotting
• ISSN: 1006-9305; 1674-7305; 1862-2798; 1869-1889
• DOI: 10.1007/s11427-007-0103-4

Plasma membrane (PM) proteome is one of the major subproteomes present in the cell, and is very important in liver function. In the present work, C57 mouse liver PM was purified by density-gradient centrifugation. The purified PM was verified by electron microscope analysis and Western blotting. The results showed that the PM was enriched by more than 20-fold and the contamination of mitochondria was reduced by 2-fold compared with the homogenization fraction. Proteins were separated by 2DE and 1DE, trypsin-digested and submitted to ESI-Q-TOF and MALDI-TOF-TOF mass spectrometry or directly digested in solution and analyzed by LC-ESI ion trap mass spectrometry. In all, 547 non-redundant mouse liver PM proteins were identified, of which 34% contributed to plasma membrane or plasma membrane-related proteins. This study optimized and evaluated the HLPP plasma membrane proteome analysis method and made a systematic analysis on PM proteome.