LysargiNase and Chemical Derivatization Based Strategy for Facilitating In-Depth Profiling of C-Terminome

doi:10.1021/acs.analchem.9b03543

	LysargiNase and Chemical Derivatization Based Strategy for Facilitating In-Depth Profiling of C-Terminome
	Hu, Hao 1; Zhao, Wensi 1,2; Zhu, Mengdi 1,2; Zhao, Lei 1; Zhai, Linhui 1; Xu, Jun-yu 1; Liu, Ping 1; Tan, Minjia 1,2
刊名	ANALYTICAL CHEMISTRY
	2019-11-19
卷号	91 期号:22 页码:14522-14529
ISSN号	0003-2700
DOI	10.1021/acs.analchem.9b03543
通讯作者	Tan, Minjia(mjtan@simm.ac.cn)
英文摘要	Global identification of protein C-termini is highly challenging due to their low abundance in conventional shotgun proteomics. Several enrichment strategies have been developed to facilitate the detection of C-terminal peptides. One major issue of previous approaches is the limited C-terminome coverage. Herein, we integrated LysargiNase digestion, chemical acetylation on neo-N-terminus, and a-ion-aided peptide matching into poly(allylamine)-based C-terminomics (termed as LAACTer). In this strategy, we leveraged LysargiNase, a protease with cleavage specificity N-terminal to Lys and Arg residues, to cover previously unidentifiable C-terminome and employed chemical acetylation and a-ion-aided peptide matching to efficiently boost peptide identifications. Triplicates of LAACTer identified a total of 834 C-termini from proteome of 293T cell, which expanded the coverage by 164% (643 more unique C-termini) compared with the parallel experiments using the original workflow. Compared with the largest human C-terminome data sets (containing 800-900 C-termini), LAACTer not only achieved comparable profiling depth but also yielded 465 previously unidentified C-termini. In a SILAC (stable isotope labeling with amino acids in cell culture)-based quantitative study for identification of GluC-cleaved products, LAACTer quantified 300% more C-terminal peptides than the original workflow. Using LAACTer and the original workflow, we performed global analysis for the C-terminal sequences of 293T cell. The original and processed C-termini displayed distinct sequence patterns, implying the "C-end rules" that regulates protein stability could be more complex than just amino acid motifs. In conclusion, we reason LAACTer could be a powerful proteomic tool for in-depth C-terminomics and would benefit better functional understanding of protein C-termini.
资助项目	National Natural Science Foundation of China[31670066] ; National Natural Science Foundation of China[91753203] ; National Natural Science Foundation of China[21907100] ; National Science and Technology Major Project Key New Drug Creation and Manufacturing Program[2018ZX09711002-004] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDA12050406] ; Shanghai Sailing Program[17YF1423200] ; Youth Innovation Promotion Association CAS
WOS关键词	PROTEIN ; PEPTIDES ; IDENTIFICATION ; PROTEOMICS ; COMPLEMENTARY ; PLASMA
WOS研究方向	Chemistry
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000498280100049
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/281988]
专题	中国科学院上海药物研究所
通讯作者	Tan, Minjia
作者单位	1.Chinese Acad Sci, Shanghai Inst Materia Med, State Key Lab Drug Res, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Hu, Hao,Zhao, Wensi,Zhu, Mengdi,et al. LysargiNase and Chemical Derivatization Based Strategy for Facilitating In-Depth Profiling of C-Terminome[J]. ANALYTICAL CHEMISTRY,2019,91(22):14522-14529.
APA	Hu, Hao.,Zhao, Wensi.,Zhu, Mengdi.,Zhao, Lei.,Zhai, Linhui.,...&Tan, Minjia.(2019).LysargiNase and Chemical Derivatization Based Strategy for Facilitating In-Depth Profiling of C-Terminome.ANALYTICAL CHEMISTRY,91(22),14522-14529.
MLA	Hu, Hao,et al."LysargiNase and Chemical Derivatization Based Strategy for Facilitating In-Depth Profiling of C-Terminome".ANALYTICAL CHEMISTRY 91.22(2019):14522-14529.