Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor

doi:10.1021/acs.analchem.9b00477

	Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor
	Lu, Yan 1,2,3; Qin, Shanshan 1; Zhang, Bingjie 1; Dai, Antao 4,5; Cai, Xiaoqing 4,5; Ma, Mengna 1,2,3; Gao, Zhan-Guo 6; Yang, Dehua 4,5; Stevens, Raymond C.1,2; Jacobson, Kenneth A.6
刊名	ANALYTICAL CHEMISTRY
	2019-07-02
卷号	91 期号:13 页码:8162-8169
ISSN号	0003-2700
DOI	10.1021/acs.analchem.9b00477
通讯作者	Wang, Ming-Wei(mwwang@simm.ac.cn) ; Shui, Wenqing(shuiwq@shanghaitech.edu.cn)
英文摘要	Affinity mass spectrometry (MS) enables rapid screening of compound mixtures for ligands bound to a specific protein target, yet its current throughput is limited to individually assay pools of 400-2000 compounds. Typical affinity MS screens implemented in pharmaceutical industry laboratories identify putative ligands based on qualitative analysis of compound binding to the target whereas no quantitative information is acquired to discriminate high- and low-affinity ligands in the screening phase. Furthermore, these screens require purification of a stabilized form of the protein target, which poses a great challenge for membrane receptor targets. Here, we describe a new, potentially general affinity MS strategy that allows screening of 20,000 compounds in one pool for highly efficient ligand discovery toward a G protein-coupled receptor (GPCR) target. Quantitative measurement of compound binding to the receptor enables high-affinity ligand selection using both the purified receptor and receptor-embedded cell membranes. This high-throughput, label-free and quantitative affinity MS screen resulted in discovery of three new antagonists of the A(2A) adenosine receptor.
资助项目	ShanghaiTech University ; National Key Research and Development Program of China[2018YFA0507004] ; National Natural Science Foundation of China[81573479] ; National Natural Science Foundation of China[81773792] ; National Mega R&D Program for Drug Discovery[2018ZX09711002-002-005] ; National Mega R&D Program for Drug Discovery[2018ZX09735-001] ; NIDDK[ZIADK31117]
WOS关键词	BINDING MODE ; IDENTIFICATION ; INHIBITORS ; DISCOVERY ; DESIGN ; ASSAYS ; GPCRS
WOS研究方向	Chemistry
语种	英语
出版者	AMER CHEMICAL SOC
WOS记录号	WOS:000474477900027
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/289323]
专题	中国科学院上海药物研究所
通讯作者	Wang, Ming-Wei; Shui, Wenqing
作者单位	1.ShanghaiTech Univ, iHuman Inst, Shanghai 201210, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Chinese Acad Sci, Natl Ctr Drug Screening, Shanghai 201203, Peoples R China 5.Chinese Acad Sci, CAS Key Lab Receptor Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China 6.NIDDK, NIH, Bethesda, MD 20892 USA 7.Fudan Univ, Sch Pharm, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Lu, Yan,Qin, Shanshan,Zhang, Bingjie,et al. Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor[J]. ANALYTICAL CHEMISTRY,2019,91(13):8162-8169.
APA	Lu, Yan.,Qin, Shanshan.,Zhang, Bingjie.,Dai, Antao.,Cai, Xiaoqing.,...&Shui, Wenqing.(2019).Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor.ANALYTICAL CHEMISTRY,91(13),8162-8169.
MLA	Lu, Yan,et al."Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor".ANALYTICAL CHEMISTRY 91.13(2019):8162-8169.