A method for isotope labeling of peptides at both ends for de novo protein sequencing
CN117890599A · Academy of Military Medical Sciences · priority 2024
| Patent | CN117890599A ↗ |
| Status | Pending |
| Priority date | 2024-01-08 |
| Filed | 2024-01-08 |
| Published | 2024-04-16 |
| Assignee | Academy of Military Medical Sciences |
Abstract
The invention discloses a method for labeling isoheavy isotopes at two ends of a peptide segment, which can be used for de novo sequencing of proteins, and can be used for de novo sequencing of peptide segments and proteins. In the method, the peptide fragment to be sequenced is divided into two parts, one part is firstly subjected to C-terminal treatment 18 O substitution, then this part of the peptide is marked with a light N-terminal acetylation, and another part of the peptide is marked with a heavy N-terminal acetylation. And after the two parts of peptide are desalted and the concentration is measured, the two parts of peptide are mixed in equal quantity for mass spectrum sample loading, and then the ion pair b with the mass difference of 2.0067Da and the ion pair y with the mass difference of 2.0017Da can be obtained, and in a high-resolution mass analyzer, the two ion pairs can be distinguished obviously. And (3) further using an algorithm to reserve one of the b/y ion pairs, removing noise and interference peaks in the spectrogram, and using the simplified spectrogram for further de-header sequencing.
Why it is here
Academy of Military Medical Sciences: labelling both peptide termini (C-terminal 18O, N-terminal 13C acetyl) for protein de novo sequencing.
Inventors (2)
秦伟捷, 陈永乐
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