[Extraction and two-dimensional liquid chromatographic fractionation of plasma membrane proteome of mouse liver cells].
OBJECTIVE
To extract the plasma membrane proteins from mouse liver cells and investigate the approach for fractionating the protein mixtures by two-dimensional liquid chromatography.


METHODS
The 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.


RESULTS
We 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.


CONCLUSIONS
This 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.
Li
Hong-Mei
HM
Department of Pathophysiology and Key Laboratory of Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, China.
Chen
Li
L
Xia
Gao-Xiao
GX
Zhao
Ming-Zhe
MZ
Hu
Shui-Wang
SW
Jiang
Yong
Y
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D008099:Liver / Q000166:cytology*; D008297:Male; D051379:Mice; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods
D020543:Proteome