Method and kit for de novo sequencing of protein N-terminal sequence by using MALDI-TOT-TOF mass spectrum
CN104483374A · Peking University · priority 2014
| Patent | CN104483374A ↗ |
| Status | Granted |
| Priority date | 2014-12-02 |
| Filed | 2014-12-02 |
| Published | 2015-04-01 |
| Assignee | Peking University |
| Also published as | CN104483374B |
Abstract
Translated from Chinese 本发明提供了一种用MALDI-TOT-TOF(基质辅助激光解吸电离-串联飞行时间)质谱对蛋白质N端序列进行从头测序的方法和试剂盒。该方法涉及蛋白质的分离、还原烷基化、蛋白质的化学修饰、酶切肽段的质谱分析、蛋白质N端肽段的质谱识别与从头测序。该方法对胶内蛋白质的赖氨酸进行胍基化，选择性地将TMPP-Ac修饰在蛋白质N端氨基上；并通过加入终止试剂和洗涤。该方法所得的蛋白质N端氨基修饰物不受过量的试剂和相应的副产物的影响，与质谱技术相容。该方法可对蛋白质的N端肽进行富集，结合反相液相色谱和生物质谱技术，既容易确定蛋白质的N端肽段，又可以很方便地进行从头测序。 The invention provides a method and a kit for de novo sequencing of protein N-terminal sequences by using MALDI-TOT-TOF (matrix-assisted laser desorption ionization-tandem time-of-flight) mass spectrometry. The method involves separation of proteins, reductive alkylation, chemical modification of proteins, mass spectrometry analysis of enzyme-cut peptides, mass spectrometry identification and de novo sequencing of protein N-terminal peptides. The method guanidinates the lysine of the protein in the gel, and selectively modifies TMPP-Ac on the N-terminal amino group of the protein; and by adding a termination reagent and washing. The protein N-terminal amino modification obtained by the method is not affected by excess reagents and corresponding by-products, and is compatible with mass spectrometry. This method can enrich the N-terminal peptides of proteins, combined with reversed-phase liquid chromatography and biological mass spectrometry, it is not only easy to determine the N-terminal peptides of proteins, but also convenient for de novo sequencing.
Why it is here
Peking University: de novo sequencing of protein N-terminal sequences by MALDI-TOF/TOF.
Inventors (2)
邹霞娟, 杨彬
An inventor links to an author page only where the same person is evident, never on a name alone.
Catalogued papers it cites (1)
- A method for N-terminal de novo sequence analysis of proteins by matrix-assisted laser desorption/ionization mass spectrometry (2008) added by the examiner
Matched from the patent’s 4 literature citations by the rules the paper citation graph uses.
Cited by (7 patent families, 6 organisations)
- 中国科学院大连化学物理研究所 academic: CN105738169A, CN109142611A
- 上海快序生物科技有限公司 company: CN116465992A
- 复旦大学 academic: CN105319116A
- 宁波大学 academic: CN119643873A
- 深圳华大智造科技股份有限公司 company: WO2023240563A1
- 詹森生物科技公司 company: CN114829927A
Every family, of any applicant, that cites this one, as Google Patents lists them; an assignee is given its English name.