High-sensitivity electron capture dissociation tandem FTICR mass spectrometry of microelectrosprayed peptides
High-sensitivity electron capture dissociation tandem FTICR mass spectrometry of microelectrosprayed peptides
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DOI：
10.1021/ac010141z
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发表时间：
2001-08-01
影响因子：
7.4
通讯作者：
Marshall, AG
中科院分区：
文献类型：
--
作者：
Håkansson, K;Emmett, MR;Marshall, AG
电子捕获解离（ECD）先前已被其他研究小组证明比其他离子解离技术产生更大的肽序列覆盖范围，并定位不稳定的翻译后修饰。在这里，ECD已经实现了10 nM （10 fmol/muL）溶液中微电喷涂的10-13聚肽和28 kda蛋白的50 nM未分离消化的色氨酸肽。串联傅立叶变换离子回旋共振（FTICR）质谱包含与所有可能的肽主胺键的裂解相对应的片段离子，除了脯氨酸的n端，对于P物质和神经紧张素。对于黄体生成素释放激素，除两种预期的主胺键断裂外，其余均观察到。该色氨酸的串联FTICR质谱包含与12个（1545.7-Da肽）中的6个和21个（2944.5-Da肽）预期主胺键中的8个对应的片段离子。目前的灵敏度比以前报道的高200-2000倍。这些结果表明，ECD有望作为一种工具，产生序列标签，用于鉴定数量有限的复杂混合物中的肽，如蛋白质组学。
Electron capture dissociation (ECD) has previously been shown by other research groups to result in greater peptide sequence coverage than other ion dissociation techniques and to localize labile posttranslational modifications. Here, ECD has been achieved for 10-13-mer peptides microelectrosprayed from 10 nM (10 fmol/muL) solutions and for tryptic peptides from a 50 nM unfractionated digest of a 28-kDa protein. Tandem Fourier transform ion cyclotron resonance (FTICR) mass spectra contain fragment ions corresponding to cleavages at all possible peptide backbone amine bonds, except on the N-terminal side of proline, for substance P and neurotensin. For luteinizing hormone-releasing hormone, all but two expected backbone amine bond cleavages are observed. The tandem FTICR mass spectra of the tryptic peptides contain fragment ions corresponding to cleavages at 6 of 12 (1545.7-Da peptide) and 8 of 21 (2944.5-Da peptide) expected backbone amine bonds. The present sensitivity is 200-2000 times higher than previously reported. These results show promise for ECD as a tool to produce sequence tags for identification of peptides in complex mixtures available only in limited amounts, as in proteomics.