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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
DOI10.1074/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.

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来源：计算技术研究所

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