公开(公告)号：US20100029005A1

公开(公告)日：2010-02-04

申请号：US12093133

申请日：2006-10-27

申请人： Junichi Kamiie ,  Sumio Otsuki ,  Tetauya Terasaki

发明人： Junichi Kamiie ,  Sumio Otsuki ,  Tetauya Terasaki

IPC分类号： G01N33/68

CPC分类号： G01N30/88 ,  G01N30/7266 ,  G01N30/8679 ,  G01N33/6848 ,  G01N2030/8813 ,  G01N2030/8818 ,  G01N2030/8827 ,  G01N2030/8831 ,  G01N2333/705 ,  H01J49/00

摘要： The present invention is to provide a method for quantifying a plasma membrane protein present in a cell membrane, by using a stable-isotope labeled peptide as a probe by mass spectrometry, in a simple, quickly and accurate manner. A plasma membrane protein is fragmented to prepare an oligopeptide fragment, identified by LC-MSMS, and by setting as essential criteria that the peptide is obtained by fragmentizing with a protease, and the peptide is specific to a target molecule, selective criteria with score are set for hydrophobic amino acids content, sequence conditions, number of amino acid residues, specific amino acid sequence conditions, etc. According to criterion for selecting a subject peptide for quantification for selecting preferentially a peptide with high score, a peptide fragment that can be ionized by ESI method is selected, and by using the subject peptide for quantification and a stable-isotope labeled peptide, the plasma membrane protein is quantified accurately by mass spectrometry.

摘要翻译： 本发明提供一种以简单，快速且准确的方式通过质谱法使用稳定同位素标记肽作为探针来定量细胞膜中存在的质膜蛋白质的方法。 将质膜蛋白质片段化以制备通过LC-MSMS鉴定的寡肽片段，并且通过设定通过用蛋白酶片段化获得肽的基本标准，并且肽对靶分子是特异性的，得分的选择性标准 设定疏水性氨基酸含量，序列条件，氨基酸残基数，特定氨基酸序列条件等。根据选择目标肽进行定量以优先选择具有高分数的肽的标准，可以离子化的肽片段 通过ESI方法，通过使用目标肽进行定量和稳定同位素标记的肽，通过质谱法精确定量质膜蛋白质。