日本プロテオーム学会大会要旨集
日本ヒトプロテオーム機構第6回大会
セッションID: P-41
会議情報

膜タンパク質をターゲットとしたプロテオーム解析基盤技術の確立
*竹中 聡松下 佳代平山 未央榊原 陽一Ming-Cheh Liu水光 正仁
著者情報
会議録・要旨集 フリー

詳細
抄録

[Introduction] Two-dimensional gel electrophoresis is a powerful technique enabling visualization of the proteome. Recently, it is reported that fluorescence labeled two-dimensional gel electrophoresis is one of the most powerful method for comprehensive proteomic analysis. While annotated two-dimensional gel electrophoresis contain thousands of proteins, they do not represent the entire genome. Hydrophobic membrane proteins in particular are conspicuously absent from such data. There are a lot of membrane proteins with important physiological functions that is involved in signal transduction etc. Recently, the improvement of the solubilize conditions for these membrane proteins are required. Therefore, we examine the application of several non-ionic detergent for two-dimensional gel electrophoresis. [Methods] Microsomal membrane fraction was obtained by centrifugation of rat brain homogenates. Membrane fraction was solubilized with sample buffer including different detergent. Two-dimensional gel electrophoresis, isoelectric focusing (IEF) in the first dimension and SDS-PAGE in the second dimension, was performed using IEF Cell (Bio-Rad). [Results] We examined some detergents and non-detergent sulfobetaines, and compared them with CHAPS that is generally used in two-dimensional gel electrophoresis. As a result, it was suggested that some non-ionic detergents improved separation in particular high molecular weight protein region. We were able to identify some membrane proteins by peptide mass fingerprinting methods using MALDI-TOF/TOF MS. This study provides methodological tools to study particular classes of membrane proteins and should be applicable to other cellular membranes such as raft microdomain.

著者関連情報
© 2008 日本プロテオーム学会(日本ヒトプロテオーム機構)
前の記事 次の記事
feedback
Top