Sequencing of Sulfopeptides Using Negative-Ion Tandem Mass Spectrometry with Hydrogen Attachment/Abstraction Dissociation
Sequencing of Sulfopeptides Using Negative-Ion Tandem Mass Spectrometry with Hydrogen Attachment/Abstraction Dissociation
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DOI：
10.1021/acs.analchem.9b01568
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发表时间：
2019-08-20
影响因子：
7.4
通讯作者：
Tanaka, Koichi
中科院分区：
文献类型：
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作者：
Asakawa, Daiki;Takahashi, Hidenori;Tanaka, Koichi
串联质谱（MS/MS）与自由基为基础的碎片，涉及连接或提取氢的肽，在一个过程中称为氢连接/提取解离（HAD），最近已经开发出来。HAD-MS/MS被认为是分析具有翻译后修饰（PTM）的蛋白质的有用方法，因为其能够确定蛋白质上的PTM位点。本研究采用负离子HAD-MS/MS结合基质辅助激光解吸/电离（MALDI）和电喷雾电离（ESI）对高酸性硫肽和硫蛋白进行分析。一般来说，MALDI和ESI分别产生单电荷和多电荷肽。HAD的单去质子化的磺肽优先产生碎片离子与磺化，而磺化和非磺化碎片离子中观察到的HAD-MS/MS光谱的多去质子化的磺肽。MALDI和ESI HAD-MS/MS谱的比较允许区分磺化和非磺化片段，这将有助于进行从头测序的磺肽。此外，基于ESI的HAD-MS/MS和液相色谱（LC）的组合允许分析存在于蛋白酶中的磺肽。HAD的LC-ESI-MS/MS是一种潜在的有用的方法用于磺基蛋白质组学的应用。
Tandem mass spectrometry (MS/MS) with radical-based fragmentation involving the attachment or abstraction of hydrogen to peptides, in a process called hydrogen attachment/abstraction dissociation (HAD), has been recently developed. HAD-MS/MS is considered a useful method for the analysis of proteins with post-translational modification (PTM) because of its ability to determine the PTM site on proteins. In the present investigation, we analyzed highly acidic sulfopeptides and sulfoprotein digests using negative-ion HAD-MS/MS combined with matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). In general, MALDI and ESI produced singly and multiply charged peptides, respectively. HAD of singly deprotonated sulfopeptides preferentially produced fragment ions with sulfonation, whereas both sulfonated and nonsulfonated fragment ions were observed in the HAD-MS/MS spectrum of multiply deprotonated sulfopeptides. A comparison of the MALDI and ESI HAD-MS/MS spectra allows the discrimination of sulfonated and nonsulfonated fragments, which would be helpful in performing de novo sequencing of sulfopeptides. In addition, the combination of ESI-based HAD-MS/MS and liquid chromatography (LC) allows the analysis of sulfopeptides present in protein digests. LC-ESI-MS/MS with HAD is a potentially useful method for sulfoproteomic application.