Extraction and two-dimensional liquid chromatographic fractionation of plasma membrane proteome of mouse liver cells / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To extract the plasma membrane proteins from mouse liver cells and investigate the approach for fractionating the protein mixtures by two-dimensional liquid chromatography.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>