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Evaluation of strategy for analyzing mouse liver plasma membrane proteome

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Abstract

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.

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Abbreviations

PM:

Plasma membrane

2DE:

two-dimensional electrophoresis

ESIQ-TOF:

electrospray ionization-quadropole-time of flight mass spectrometry

MALDI-TOF:

matrix-assisted laser-desorption/ionization time of flight

LC-MS/MS:

liquid chromatography coupled with tandem mass spectrometry

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Authors and Affiliations

  1. College of Life Sciences, Hunan Normal University, Changsha, 410081, China

    Chen Ping, Zhang LiJun, Li XuanWen, Wang Xie, Cao Rui, Liu Zhen, Xiong JiXian, Peng Xia, Wei YingJuan, Ying XingFeng, Wang XianChun & Liang SongPing

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  1. Chen Ping
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  2. Zhang LiJun
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  3. Li XuanWen
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  4. Wang Xie
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  5. Cao Rui
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  7. Xiong JiXian
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  8. Peng Xia
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Corresponding author

Correspondence to Liang SongPing.

Additional information

These authors contributed equally to this work

Supported by China National Haman Liver Proteome Project (Grant No. 2004 BA711A18), and Hunan Provincial Education Department and Hunan Science and Technology Project (Grant Nos. 05FJ2002 and 05FJ4018)

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Chen, P., Zhang, L., Li, X. et al. Evaluation of strategy for analyzing mouse liver plasma membrane proteome. SCI CHINA SER C 50, 731–738 (2007). https://doi.org/10.1007/s11427-007-0103-4

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  • DOI: https://doi.org/10.1007/s11427-007-0103-4

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