小鼠肝脏细胞质膜蛋白质组的提取和二维液相色谱分离  

Extraction and two-dimensional liquid chromatographic fractionation of plasma membrane proteome of mouse liver cells

作　　者：李红梅[1] 陈丽[1] 夏高晓[1] 赵明哲[1] 胡水旺[1] 姜勇[1] 

机构地区：[1]南方医科大学病理生理学教研室和广东省蛋白质组学重点实验室,广东广州510515

出　　处：《南方医科大学学报》2008年第11期1947-1949,1953,共4页Journal of Southern Medical University

基　　金：国家自然科学基金委-广东省人民政府自然科学联合基金重点项目(U0632004);国家自然科学基金(30572151;30670828);教育部长江学者和创新团队发展计划项目(IRT0731)

摘　　要：目的提取小鼠肝脏细胞质膜蛋白并建立一种利用二维液相色谱法分离细胞质膜蛋白质组的方法。方法采用差速离心结合蔗糖密度梯度离心分离和富集分小鼠肝脏细胞质膜蛋白,样品经脱盐及用起始缓冲液置换后,进行一维色谱聚焦分离,然后收集pH8.5～4.0之间的组分进行二维反相高效液相色谱分离,最后将获得的二维UV图通过ProteoVue软件转换成UV/pI图谱。结果成功提取了肝脏细胞质膜蛋白,并通过二维液相色谱成功建立了小鼠肝脏细胞质膜蛋白的二维UV/pI图谱,收集了一维色谱聚焦分离的pH8.5～4.0区间的16个组分,并将每个组分分进行二维色谱分离后转换为UV/pI图谱。结论为进一步全面研究小鼠肝脏细胞质膜蛋白功能和疾病差异蛋白质组研究打下了基础。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. Conclusion 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.

关 键 词：细胞质膜蛋白质 二维液相色谱 蛋白质组 肝脏 

分 类 号：Q51-3[生物学—生物化学]