C-terminal peptide sequencing using acetylated peptides with MSn in a quadrupole ion trap.
C-terminal peptide sequencing using acetylated peptides with MSn in a quadrupole ion trap.
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在四极离子阱中使用乙酰化肽和 MSn 进行 C 端肽测序。
DOI：
10.1039/a908950k
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Glish,GL
中科院分区：
文献类型：
--
作者：
Payne,AH;Chelf,JH;Glish,GL
质谱/质谱法用于多肽和小蛋白的测序已有多年历史。这种方法可以分离出感兴趣的肽，从而可以分析不纯的样品和未分离的混合物，例如蛋白质消化。所选肽离子的碰撞诱导解离（CID）产生提供序列信息的产物离子。然而，通常MS/MS谱不能提供足够的信息来确定完整的序列。四极离子阱具有多级质谱、MSn的功能，增加了确定肽序列的信息。规则和可预测的多肽解离模式进一步简化了这一分析。通过主要形成一种类型的离子，消除了离子是N端还是c端的模糊性。这种模式也有利于离子强度在质谱/质谱的下一阶段保持集中。在这项工作中，探索了一种通过控制解离途径来利用四极离子阱的MSn能力的方法。通过使肽的n端乙酰化来改变解离。利用各种乙酰化肽的MSn来确定c端残基的身份和肽的长度对所观察到的解离途径的影响。
MS/MS has been used to sequence peptides and small proteins for a number of years. This method allows one to isolate the peptide of interest, which makes it possible to analyze impure samples and unseparated mixtures, such as protein digests. Collision-induced dissociation (CID) of the selected peptide ion generates the product ions that provide sequence information. However, often the MS/MS spectrum does not provide adequate information for complete sequence determination. The quadrupole ion trap has the capability to do multiple stages of mass spectrometry, MSn, which can increase the information available to determine the peptide sequence. A regular and predictable dissociation pattern for peptides further simplifies this analysis. By forming predominantly one type of ion, ambiguity is removed as to whether the ion is N- or C-terminal. This pattern can also be advantageous in that ion intensity remains concentrated for the next stage of MS/MS. In this work, a method to take advantage of the MSn capabilities of the quadrupole ion trap by controlling the dissociation pathways is explored. Dissociation is altered by acetylating the N-terminus of the peptide. MSn of a variety of acetylated peptides is used to determine the effects of the identity of the C-terminal residue and the length of the peptide on the dissociation pathways observed.