Extraction and two-dimensional liquid chromatographic fractionation of plasma membrane proteome of mouse liver cells.
- Author:
Hong-Mei LI
1
;
Li CHEN
;
Gao-Xiao XIA
;
Ming-Zhe ZHAO
;
Shui-Wang HU
;
Yong JIANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Membrane; metabolism; Chemical Fractionation; instrumentation; methods; Chromatography, Liquid; methods; Liver; cytology; Male; Mice; Proteome; metabolism; Proteomics; instrumentation; methods
- From: Journal of Southern Medical University 2008;28(11):1947-1949
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo extract the plasma membrane proteins from mouse liver cells and investigate the approach for fractionating the protein mixtures by two-dimensional liquid chromatography.
METHODSThe plasma membrane of the liver cells from 10 mice was extracted by differential centrifugation and sucrose density-gradient centrifugation. The plasma membrane proteins were exchanged with the start buffer and separated by chromatofocusing in the first-dimensional fractionation. The final results were transformed into UV/pI maps using ProteoVue software.
RESULTSWe successfully extracted the plasma membrane proteins from mouse liver cells. Sixteen fractions between pH 8.5-4.0 were recovered in the first-dimensional chromatofocusing followed by 2D- chromatographic fractionation, and the results were displayed as UV/pI maps.
CONCLUSIONThis approach for fractionating the mouse liver cell plasma membrane protein study provides the foundation for further studies on the functions of plasma membrane proteins and differential proteome of diseases.