Precision De Novo Peptide Sequencing Using Mirror Proteases of Ac-LysargiNase and Trypsin for Large-scale Proteomics
文献类型:期刊论文
作者 | Wu, Fei-Lin4,5,6,7; Zengt, Wen-Feng8,9,10; Overall, Christopher M.2; Chi, Hao8,9,10; He, Si-Min8,9,10; Wang, Fu-Qiang4,5,6,7; Li, Yan-Chang4,5,6,7; Xu, Feng4,5,6,7; Zhao, Ming-Zhi4,5,6,7; Wang, Xi8,9,10 |
刊名 | MOLECULAR & CELLULAR PROTEOMICS
![]() |
出版日期 | 2019-04-01 |
卷号 | 18期号:4页码:773-785 |
ISSN号 | 1535-9476 |
DOI | 10.1074/mcp.TIR118.000918 |
英文摘要 | De novo peptide sequencing for large-scale proteomics remains challenging because of the lack of full coverage of ion series in tandem mass spectra. We developed a mirror protease of trypsin, acetylated LysargiNase (Ac-LysargiNase), with superior activity and stability. The mirror spectrum pairs derived from the Ac-LysargiNase and trypsin treated samples can generate full b and y ion series, which provide mutual complementarity of each other, and allow us to develop a novel algorithm, pNovoM, for de novo sequencing. Using pNovoM to sequence peptides of purified proteins, the accuracy of the sequence was close to 100%. More importantly, from a large-scale yeast proteome sample digested with trypsin and Ac-LysargiNase individually, 48% of all tandem mass spectra formed mirror spectrum pairs, 97% of which contained full coverage of ion series, resulting in precision de novo sequencing of full-length peptides by pNovoM. This enabled pNovoM to successfully sequence 21,249 peptides from 3,753 proteins and interpreted 44-152% more spectra than pNovo+ and PEAKS at a 5% FDR at the spectrum level. Moreover, the mirror protease strategy had an obvious advantage in sequencing long peptides. We believe that the combination of mirror protease strategy and pNovoM will be an effective approach for precision de novo sequencing on both single proteins and proteome samples. |
资助项目 | National Key R&D Program of China[2016YFA0501300] ; National Key R&D Program of China[2017YFC0906600] ; National Key R&D Program of China[2017YFA0505002] ; National Key R&D Program of China[2015CB910700] ; National Natural Science Foundation of China[31870824] ; National Natural Science Foundation of China[91839302] ; National Natural Science Foundation of China[31670834] ; National Natural Science Foundation of China[31700723] ; Youth Innovation Promotion Association CAS[2014091] ; National Megaprojects for Key Infectious Diseases[2018ZX10302302001] ; National Natural Science Foundation of Beijing[5152008] ; Beijing Training Project for The Leading Talents in ST[Z161100004916024] ; Foundation of State Key Laboratory of Proteomics[SKLP-Y201501] ; Foundation of State Key Laboratory of Proteomics[SKLP-K201705] ; Innovation Foundation of Medicine[2017CXJJ19] ; Innovation Foundation of Medicine[BWS17J032] ; Innovation Foundation of Medicine[16CXZ027] ; Innovation Foundation of Medicine[BWS14J052] ; CAS Interdisciplinary Innovation Team[Y604061000] |
WOS研究方向 | Biochemistry & Molecular Biology |
语种 | 英语 |
WOS记录号 | WOS:000466934700012 |
出版者 | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC |
源URL | [http://119.78.100.204/handle/2XEOYT63/4245] ![]() |
专题 | 中国科学院计算技术研究所期刊论文_英文 |
通讯作者 | Chi, Hao; He, Si-Min; Xu, Ping |
作者单位 | 1.Hebei Univ, Coll Life Sci, Baoding 071002, Peoples R China 2.Univ British Columbia, Ctr Blood Res, Vancouver, BC, Canada 3.Wuhan Univ, Minist Educ, Key Lab Combinatorial Biosynth & Drug Discovery, Sch Pharmaceut Sci, Wuhan 430071, Hubei, Peoples R China 4.Beijing Beijing Inst Life, Beijing 102206, Peoples R China 5.Natl Ctr Prot Sci Beijing, Beijing 102206, Peoples R China 6.Beijing Proteome Res Ctr, Beijing 102206, Peoples R China 7.State Key Lab Prote, Beijing 102206, Peoples R China 8.Chinese Acad Sci, Inst Comp Technol, Beijing 100190, Peoples R China 9.Univ Chinese Acad Sci, Beijing 100190, Peoples R China 10.Chinese Acad Sci, Inst Comp Technol, Key Lab Intelligent Informat Proc, Beijing 100190, Peoples R China |
推荐引用方式 GB/T 7714 | Wu, Fei-Lin,Zengt, Wen-Feng,Overall, Christopher M.,et al. Precision De Novo Peptide Sequencing Using Mirror Proteases of Ac-LysargiNase and Trypsin for Large-scale Proteomics[J]. MOLECULAR & CELLULAR PROTEOMICS,2019,18(4):773-785. |
APA | Wu, Fei-Lin.,Zengt, Wen-Feng.,Overall, Christopher M..,Chi, Hao.,He, Si-Min.,...&Yang, Hao.(2019).Precision De Novo Peptide Sequencing Using Mirror Proteases of Ac-LysargiNase and Trypsin for Large-scale Proteomics.MOLECULAR & CELLULAR PROTEOMICS,18(4),773-785. |
MLA | Wu, Fei-Lin,et al."Precision De Novo Peptide Sequencing Using Mirror Proteases of Ac-LysargiNase and Trypsin for Large-scale Proteomics".MOLECULAR & CELLULAR PROTEOMICS 18.4(2019):773-785. |
入库方式: OAI收割
来源:计算技术研究所
浏览0
下载0
收藏0
其他版本
除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。