A de novo sequencing method by mass spectrometry based on selective enzyme labeling of the C-terminus of peptides
CN109900834B · Dalian Institute of Chemical Physics · priority 2017
| Patent | CN109900834B ↗ |
| Status | Active |
| Priority date | 2017-12-07 |
| Filed | 2017-12-07 |
| Published (granted) | 2022-05-17 |
| Assignee | Dalian Institute of Chemical Physics |
| Also published as | CN109900834A |
Abstract
Translated from Chinese 本发明涉及一种基于肽段C末端选择性酶标记的质谱从头测序方法及其应用。所述的方法是在肽段C末端酰胺酶的催化作用下，肽段C末端的羧基直接通过酰胺化反应标记上带碱性基团的小分子或者先通过酰胺化反应标记上带氨基的不饱和烃类，进一步通过叠氮‑炔基或者巯基‑烯的点击化学反应连接上碱性功能分子。肽段C末端标记上的碱性分子增加二级质谱中y离子的数目，提高肽段在质谱中的响应和碎裂效率，从而实现肽段测序。 The invention relates to a mass spectrometry de novo sequencing method based on the selective enzyme labeling of the C-terminal of a peptide segment and its application. The method is that under the catalysis of the C-terminal amidase of the peptide segment, the carboxyl group at the C-terminal of the peptide segment is directly labeled with a small molecule with a basic group through an amidation reaction, or a small molecule with an amino group is first labeled with an amino group through an amidation reaction. Saturated hydrocarbons are further connected to basic functional molecules through the click chemistry reaction of azide-alkynyl or mercapto-ene. The basic molecules on the C-terminal label of the peptide segment increase the number of y ions in the secondary mass spectrometer, improve the response and fragmentation efficiency of the peptide segment in the mass spectrometer, and thus realize the sequence of the peptide segment.
Why it is here
Dalian Institute of Chemical Physics: de novo sequencing after enzymatic C-terminal labelling of peptides with a basic group, which strengthens the y-ion series.
Inventors (6)
张丽华, 吴琼, 单亦初, 杨超, 杨开广, 张玉奎
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