Quantification of proteins using 13C7-labeled and unlabeled iodoacetanilide by nano liquid chromatography/nanoelectrospray ionization and by selected reaction monitoring mass spectrometry
Quantification of proteins using 13C7-labeled and unlabeled iodoacetanilide by nano liquid chromatography/nanoelectrospray ionization and by selected reaction monitoring mass spectrometry
复制标题
使用 13C7 标记和未标记的碘乙酰苯胺，通过纳液相色谱/纳电喷雾电离和选择反应监测质谱法对蛋白质进行定量
DOI：
10.1016/j.bmcl.2014.12.090
复制
发表时间：
2015
期刊：
影响因子：
--
通讯作者：
S.
中科院分区：
文献类型：
--
作者：
Kurono;S.;Kaneko;Y.;Matsuura;S.;Niwayama;S.
半胱氨酸特异性修饰剂碘代乙酰苯胺（IAA）和13 C7-碘代乙酰苯胺（13 C7-IAA）的组合已被应用于蛋白质的绝对定量。选择反应监测（SRM）与纳米液相色谱/纳米电喷雾离子阱质谱（nano LC/nano-ESI-IT-MS）分析的使用被应用于三个商业蛋白质的精确定量。在简单的缓冲溶液和复杂的蛋白质环境中，所有这些蛋白质的理论比率和观察到的比率之间观察到良好的相关性。由于有效的标记，该方法不需要单独合成用于SRM研究的同位素标记的肽。因此，该方法有望成为蛋白质组学研究的有用工具。
The combination of cysteine-specific modifiers, iodoacetanilide (IAA) and13C7-labeled iodoacetanilide (13C7-IAA), has been applied to absolute quantification of proteins. The selected reaction monitoring (SRM) with the use of nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry (nano LC/nano-ESI-IT-MS) analysis was applied to precise quantification of three commercial proteins. Good correlation was observed between the theoretical ratios and observed ratios for all these proteins both in a simple buffer solution and in a complex protein environment. Due to efficient tagging, this method does not require separate synthesis of isotope-labeled peptides for the SRM studies. Therefore, this method is expected to be a useful tool for proteomics research.