Evaluation of strategy for analyzing mouse liver plasma membrane proteome

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...

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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
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cited_by cdi_FETCH-LOGICAL-c387t-ac398317b858dc08957e072246f5fad1a27ccf4c3c347c2b6069a399f3c9a4883
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creator Chen, Ping
Zhang, LiJun
Li, XuanWen
Wang, XiE
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Liu, Zhen
Xiong, JiXian
Peng, Xia
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Ying, XingFeng
Wang, XianChun
Liang, SongPing
description 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.
doi_str_mv 10.1007/s11427-007-0103-4
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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
title Evaluation of strategy for analyzing mouse liver plasma membrane proteome
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