Ujihara Tetsuro, Sakurai Isamu, Mizusawa Naoki, Wada Hajime
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Meguro-ku, Tokyo 153-8902, Japan.
Anal Biochem. 2008 Mar 15;374(2):429-31. doi: 10.1016/j.ab.2007.11.014. Epub 2007 Nov 17.
In protein analysis using mass spectrometry, proteins are usually separated by electrophoresis and digested within the gel with proteases such as trypsin. However, analysis of lipid-modified proteins is difficult due to the low recovery of lipid-modified peptide fragments from the gel as well as their low ionization efficiency during mass spectrometry. In this study, we developed a simple extraction method with n-dodecyl-beta-D-maltoside following chloroform/methanol extraction that efficiently elutes lipid-modified fragments from gels. This method allowed us to analyze the structure of lipid-modified fragments, suggesting the applicability of the method for analysis of lipid-modified fragments by mass spectrometry.
在使用质谱进行蛋白质分析时,蛋白质通常通过电泳分离,然后在凝胶中用诸如胰蛋白酶等蛋白酶进行消化。然而,由于脂质修饰的肽片段从凝胶中的回收率低以及它们在质谱分析期间的低电离效率,脂质修饰蛋白质的分析很困难。在本研究中,我们开发了一种在氯仿/甲醇提取之后用正十二烷基-β-D-麦芽糖苷的简单提取方法,该方法能有效地从凝胶中洗脱脂质修饰的片段。该方法使我们能够分析脂质修饰片段的结构,表明该方法适用于通过质谱分析脂质修饰片段。