Evaluation of the Application of Sodium Deoxycholate to Proteomic Analysis of Rat Hippocampal Plasma Membrane

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• 作者：Jian Zhou ; Tieyang Zhou ; Rui Cao ; Zhen Liu ; Jianying Shen ; Ping Chen ; Xianchun Wang ; Songping Liang
• 刊名：Journal of Proteome Research
• 出版年：2006
• 出版时间：October 2006
• 年：2006
• 卷：5
• 期：10
• 页码：2547 - 2553
• 全文大小：112K
• 年卷期：v.5,no.10(October 2006)
• ISSN：1535-3907

文摘

Detergents have been widely used for the solubilization of membrane proteins and the improvementof their digestion. In this paper, we have evaluated the application of sodium deoxycholate (SDC) tothe solubilization and digestion of rat hippocampal plasma membrane (PM) proteins. For in-solutiondigestion, rat hippocampal PM fraction from sucrose-density gradient centrifugation was solubilizedby boiling in 1.0% SDC, and directly digested without dilution. During the in-gel digestion of thehippocampal PM proteins separated by SDS-PAGE, 0.1% SDC was added. Before analysis of peptidemixture by liquid chromatography and electrospray mass spectrometry, SDC in the tryptic digests wasremoved by centrifugation following acidification. Use of 1.0% SDC in solubilization and in-solutiondigestion of rat PM proteins had led to 77 PM or membrane-associated proteins identified, a morethan 2-fold increase over that by use of SDS. The addition of 0.1% SDC to the in-gel digestion of SDS-PAGE-resolved membrane proteins remarkably enhanced the coverage of tryptic peptides and thenumber of hydrophobic membrane proteins identified. Being a cheaper and more tractable acid-insoluble detergent, SDC could be used at higher concentration in the solubilization and tryptic digestionof proteins including PM proteins with the purpose of enhancing the protein solubility and at the sametime making no interference with trypsin activity and subsequent analyses.