De novo sequencing of protein amino acid sequences based on non-isobaric markers at both ends
CN105301119B · Dalian Institute of Chemical Physics · priority 2014
| Patent | CN105301119B ↗ |
| Status | Active |
| Priority date | 2014-07-15 |
| Filed | 2014-07-15 |
| Published (granted) | 2017-06-06 |
| Assignee | Dalian Institute of Chemical Physics |
| Also published as | CN105301119A |
Abstract
The present invention relates to a kind of protein amino acid sequence de novo sequencing method based on the non-grade weight cold labeling in polypeptide two ends.Digestion is carried out to the protein after denaturation reductive alkylation with intracellular protein enzyme lysine C, the peptide fragment containing N-terminal α amino and C-terminal lysine side-chain ε amino is formed.After sample is divided into two parts, selective derivatization is carried out to polypeptide N-terminal α amino using the labelled reagent containing different isotopes；Reuse the labelled reagent containing different isotopes to derive or in cultured cells peptide C end lysine ε amino, the mark to lysine in protein is realized using the nutrient solution containing isotope marks lysine.Finally, sample is mixed, is separated and identified using high performance liquid chromatography mass spectrum.The present invention is differentiated by the peptide fragment ion for marking formation to exist in pairs, reduces the influence of interference signal, is improved fragment ion and is selected specificity and the sequencing polypeptides degree of accuracy；Improve the speed of de novo sequencing.
Why it is here
Dalian Institute of Chemical Physics: protein de novo sequencing with non-isobaric light/heavy labels at both peptide termini, which tell b ions from y ions.
Inventors (5)
张丽华, 单亦初, 张珅, 吴琪, 张玉奎
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Cited by catalogued patents (1)
- WO2025156912A1 Direct-read precision de novo sequencing method for proteins and proteome and based on product ion type identification and full product-ion coverage
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