One-Step SH2 Superbinder-Based Approach for Sensitive Analysis of Tyrosine Phosphoproteome

doi:10.1021/acs.jproteome.9b00045

	One-Step SH2 Superbinder-Based Approach for Sensitive Analysis of Tyrosine Phosphoproteome
	Li, Yanan 2,3; Yao, Yating 2,3; Wang, Yan 2,3; Wang, Shujuan 1; Liu, Xiaoyan 2,3; Liu, Zhen 2,3; Fang, Zheng 2,3; Mao, Jiawei 2,3; Zheng, Yong 1; Ye, Mingliang 2,3
刊名	JOURNAL OF PROTEOME RESEARCH
	2019-04-01
卷号	18 期号:4 页码:1870-1879
关键词	phosphoproteomics SH2 superbinder protein tyrosine phosphorylation phosphopeptide enrichment LC-MS/MS
ISSN号	1535-3893
DOI	10.1021/acs.jproteome.9b00045
通讯作者	Zheng, Yong(yong99@hotmail.com) ; Ye, Mingliang(mingliang@dicp.ac.cn)
英文摘要	Tyrosine phosphorylation plays a major role in regulating cell signaling pathways governing diverse biological functions such as proliferation and differentiation. Systemically mapping phosphotyrosine (pTyr) sites is the key to understanding molecular mechanisms underlining pTyr-dependent signaling. Although mass spectrometry-based technologies have been widely used for pTyr site profiling and quantification, their applications are often hindered by the poor efficiency in current multistep enrichment procedures for inherently low abundance pTyr peptides, especially under physiological conditions. Taking advantage of the sequence-independent high affinity of SH2 superbinder toward pTyr residues, we have developed a simplified one-step pTyr peptide enrichment method that uses immobilized SH2 superbinder for unbiased and robust enrichment of endogenous pTyr peptides from biological samples. By eliminating the prerequisite global phosphopeptide enrichment step in our previously developed two-step method, we minimized sample loss and improved peptide capture efficiency. Applying this method to Jurkat cells at resting state, where the tyrosine phosphorylation level is low, both the number of identified pTyr peptides and sites are increased by three folds compared to the two-step method. Specifically, we were able to identify 511 nonredundant pTyr peptides, corresponding to 403 high confidence pTyr sites, from Jurkat cells with high level technical reproducibility (Pearson's correlation coefficient as high as 0.94). Further applying this method to two human breast cancer cell lines, BT474 and HCC1954, before and after EGF stimulation, we demonstrated that this approach could be a powerful tool for illustrating pTyr-dependent signaling network controlling cellular behaviors such as drug resistance.
资助项目	National Key R&D Program of China[2016YFA0501402] ; National Key R&D Program of China[2017YFA0505004] ; National Key R&D Program of China[2016YFA0501404] ; National Natural Science Foundation of China[21535008] ; National Natural Science Foundation of China[21605140] ; National Natural Science Foundation of China[91753105] ; National Natural Science Foundation of China[31670839] ; the innovation program of science and research from the DICP, CAS[DICP TMSR201601] ; Natural Science Foundation of Beijing[5172012] ; National Natural Science Fund of China for Distinguished Young Scholars[21525524]
WOS关键词	TRASTUZUMAB RESISTANCE ; SIGNALING NETWORKS ; CANCER ; PHOSPHORYLATION ; BREAST ; CHROMATOGRAPHY ; PHOSPHATASE ; REVEALS ; KINASES ; SHP2
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000464068900035
资助机构	National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; the innovation program of science and research from the DICP, CAS ; the innovation program of science and research from the DICP, CAS ; Natural Science Foundation of Beijing ; Natural Science Foundation of Beijing ; National Natural Science Fund of China for Distinguished Young Scholars ; National Natural Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; the innovation program of science and research from the DICP, CAS ; the innovation program of science and research from the DICP, CAS ; Natural Science Foundation of Beijing ; Natural Science Foundation of Beijing ; National Natural Science Fund of China for Distinguished Young Scholars ; National Natural Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; the innovation program of science and research from the DICP, CAS ; the innovation program of science and research from the DICP, CAS ; Natural Science Foundation of Beijing ; Natural Science Foundation of Beijing ; National Natural Science Fund of China for Distinguished Young Scholars ; National Natural Science Fund of China for Distinguished Young Scholars ; National Key R&D Program of China ; National Key R&D Program of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; the innovation program of science and research from the DICP, CAS ; the innovation program of science and research from the DICP, CAS ; Natural Science Foundation of Beijing ; Natural Science Foundation of Beijing ; National Natural Science Fund of China for Distinguished Young Scholars ; National Natural Science Fund of China for Distinguished Young Scholars
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/165596]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Zheng, Yong; Ye, Mingliang
作者单位	1.Beijing Inst Life, Natl Ctr Prot Sci Beijing, Beijing Proteome Res Ctr, State Key Lab Prote, Beijing 102206, Peoples R China 2.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Li, Yanan,Yao, Yating,Wang, Yan,et al. One-Step SH2 Superbinder-Based Approach for Sensitive Analysis of Tyrosine Phosphoproteome[J]. JOURNAL OF PROTEOME RESEARCH,2019,18(4):1870-1879.
APA	Li, Yanan.,Yao, Yating.,Wang, Yan.,Wang, Shujuan.,Liu, Xiaoyan.,...&Ye, Mingliang.(2019).One-Step SH2 Superbinder-Based Approach for Sensitive Analysis of Tyrosine Phosphoproteome.JOURNAL OF PROTEOME RESEARCH,18(4),1870-1879.
MLA	Li, Yanan,et al."One-Step SH2 Superbinder-Based Approach for Sensitive Analysis of Tyrosine Phosphoproteome".JOURNAL OF PROTEOME RESEARCH 18.4(2019):1870-1879.