Proteomic Analysis of a Membrane Skeleton Fraction from Human Liver

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• 作者：Jintang He ; Yashu Liu ; Sizhi He ; Qingsong Wang ; Hai Pu ; Jianguo Ji
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：September 2007
• 年：2007
• 卷：6
• 期：9
• 页码：3509 - 3518
• 全文大小：699K
• 年卷期：v.6,no.9(September 2007)
• ISSN：1535-3907

文摘

The cytoskeleton networks around liver cell cortex can resist Triton extraction and co-pellet with theirtightly associated integral membrane proteins, forming assemblies called "membrane skeletons".Despite their important roles in determining cell shape and in signal transduction pathways, themembrane skeletons of human liver cells are uncharacterized to a great extent. In the present work,we prepared a membrane skeleton fraction by Triton extraction of human liver plasma membranesand then separated its protein components by 2-D gels. We optimized the detergent used for proteinsolubilization and found that 2% ASB-14 allowed the best recovery of membrane skeleton proteins. Byanalyzing the protein spots with MALDI-TOF and MALDI-TOF-TOF MS, we identified 104 nonredundantproteins, wherein 38 were cytoskeletal proteins that were further classified into several groups, includingproteins in fodrin-based meshworks, adhesion proteins (proteins involved in adherens junctions, focaladhesions, desmosomes, hemidesmosomes and tight junctions), proteins that regulate F-actin dynamics,motor proteins, and some other cytoskeletal proteins. To the best of our knowledge, this is one of thelargest data sets of membrane skeleton proteins to date. All the results suggested that the liver cellshad complex actin- and cytokeratin-based membrane skeletons. This work provided a representative2-DE map of membrane skeletons from human normal liver, for the purpose of helping to elucidatethe composition and function of the membrane skeletons.