Document Detail


Integration of a two-phase partition method into proteomics research on rat liver plasma membrane proteins.
MedLine Citation:
PMID:  16512679     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To comprehensively identify proteins of the rat liver plasma membrane (PM), we have adopted a proteomics strategy that utilizes sucrose density centrifugation in conjunction with aqueous two-phase partition for plasma membrane isolation, followed by SDS-PAGE, mass spectrometry and bioinformatics. Western blot analysis showed that this method results in highly purified plasma membrane fractions, which is a key to successful plasma membrane proteomics. The PM proteins were separated by SDS-PAGE and digested with trypsin. Through nano-ESI-LC MS/MS analysis we identified 428 rat liver membrane proteins, of which 304 had a gene ontology (GO) annotation indicating a cellular component, and 204 (67%) of the latter were known integral membrane proteins or membrane-associated proteins. In addition to proteins known to be associated with the plasma membrane, several hypothetical proteins have also been identified. This study not only provides a tool to study plasma membrane proteins with low levels of contamination, but also provides a data set for proteins of high to moderate abundance in rat liver plasma membranes, thus allowing for more comprehensive characterization of membrane proteins and a better understanding of membrane dynamics.
Authors:
Rui Cao; Xuanwen Li; Zhen Liu; Xia Peng; Weijun Hu; Xianchun Wang; Ping Chen; Jingyun Xie; Songping Liang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of proteome research     Volume:  5     ISSN:  1535-3893     ISO Abbreviation:  J. Proteome Res.     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-03-03     Completed Date:  2007-03-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101128775     Medline TA:  J Proteome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  634-42     Citation Subset:  IM    
Affiliation:
Key Laboratory of Protein Chemistry and Developmental Biology of Education Committee, College of Life Sciences, Hunan Normal University, Changsha, People's Republic of China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Membrane / chemistry,  enzymology,  metabolism*
Gene Expression Regulation
Liver / enzymology,  metabolism*
Membrane Proteins / chemistry,  isolation & purification,  metabolism*
Proteomics / methods*
Rats
Signal Transduction / physiology
Chemical
Reg. No./Substance:
0/Membrane Proteins

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