Improving de novo sequencing of peptides using a charged tag and C-terminal digestion
Improving de novo sequencing of peptides using a charged tag and C-terminal digestion
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DOI：
10.1021/ac061670b
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发表时间：
2007-02-15
影响因子：
7.4
通讯作者：
Gebler, John C.
中科院分区：
文献类型：
--
作者：
Chen, Weibin;Lee, Peter J.;Gebler, John C.
提出了一种改进的MALDI质谱多肽从头测序方法。该方法将电荷衍生化反应与c端消化相结合来修饰色氨酸肽。电荷衍生化将固定的电荷基团附着在肽的n端，衍生化步骤后的酶切去除c端碱性氨基酸残基，如精氨酸和赖氨酸。在低能CID条件下（MALDI Q-TOF质谱仪）对修饰肽进行碎片化，得到简化但完整的肽序列离子序列。该方法的有效性通过几个模型蛋白质消化的结果得到了证明，其中肽序列无论是手动还是通过自动测序程序都得到了正确的推断。
An improved method for peptide de novo sequencing by MALDI mass spectrometry is presented. The method couples a charge derivatization reaction with C-terminal digestion to modify tryptic peptides. The charge derivatization attaches a fixed charge group onto the N-termini of peptides, and the enzymatic digestion after the derivatization step removes C-terminal basic amino acid residues such as arginine and lysine. The fragmentation of the modified peptide(s) under low-energy CID conditions (MALDI Q-TOF mass spectrometer) yields a simplified yet complete ion series of the peptide sequence. The validity of the method is demonstrated by the results from several model protein digests, where peptide sequences were correctly deduced either manually or through an automated sequencing program.