Solubilization, purification, and ligand binding characterization of G protein-coupled receptor SMO in native membrane bilayer using styrene maleic acid copolymer

doi:10.7717/peerj.13381

	Solubilization, purification, and ligand binding characterization of G protein-coupled receptor SMO in native membrane bilayer using styrene maleic acid copolymer
	Zhu, Lina 1,4; Zhao, Hongxin 1; Wang, Yizhuo 1,3; Yu, Chuandi 1,3; Liu, Juanjuan 1; Li, Ling 1,3; Li, Zehua 1,3; Zhang, Jin 2; Dai, Han 1; Wang, Junfeng 1,3,4
刊名	PEERJ
	2022-05-03
卷号	10
关键词	GPCR Dissociation constant Detergents Styrene maleic acid copolymer nanodiscs Smoothened receptors NMR
ISSN号	2167-8359
DOI	10.7717/peerj.13381
通讯作者	Wang, Junfeng(junfeng@hmfl.ac.cn) ; Zhu, Lei(zhulei@hmfl.ac.cn)
英文摘要	Smoothened (SMO) protein is a member of the G protein-coupled receptor (GPCR) family that is involved in the Hedgehog (Hh) signaling pathway. It is a putative target for treating various cancers, including medulloblastoma and basal cell carcinoma (BCC). Characterizing membrane proteins such as SMO in their native state is highly beneficial for the development of effective pharmaceutical drugs, as their structures and functions are retained to the highest extent in this state. Therefore, although SMO protein is conventionally solubilized in detergent micelles, incorporating the protein in a lipid-based membrane mimic is still required. In this study, we used styrene maleic acid (SMA) copolymer that directly extracted membrane protein and surrounding lipids as well as formed the so-called polymer nanodiscs, to solubilize and purify the SMO transmembrane domain encapsulated by SMA-nanodiscs. The obtained SMA-nanodiscs showed high homogeneity and maintained the physiological activity of SMO protein, thereby enabling the measurement of the dissociation constant (K-d) for SMO ligands SMO-ligands Shh Signaling Antagonist V (SANT-1) and Smoothened Agonist (SAG) using ligand-based solution nuclear magnetic resonance spectroscopy. This work paves the way for investigating the structure, function, and drug development of SMO proteins in a native-like lipid environment.
资助项目	Foundation of Anhui Technologies Major Program[201903a07020026] ; Foundation of President of the Chinese Academy of Sciences[YZJJZX202004] ; National Natural Science Foundation of China[32171226] ; High Magnetic Field Laboratory of Anhui Province
WOS关键词	MONOMERIC INTERLEUKIN-8 ; NANOPARTICLES ; NANODISCS ; RECONSTITUTION ; SIZE ; CHANNEL ; COMPLEX ; SMALPS ; CXCR1
WOS研究方向	Science & Technology - Other Topics
语种	英语
出版者	PEERJ INC
WOS记录号	WOS:000797871100004
资助机构	Foundation of Anhui Technologies Major Program ; Foundation of President of the Chinese Academy of Sciences ; National Natural Science Foundation of China ; High Magnetic Field Laboratory of Anhui Province
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/130961]
专题	中国科学院合肥物质科学研究院
通讯作者	Wang, Junfeng; Zhu, Lei
作者单位	1.Chinese Acad Sci, Hefei Inst Phys Sci, High Magnet Field Lab, Key Lab High Magnet Field & Ion Beam Phys Biol, Hefei, Peoples R China 2.Nanchang Univ, Sch Basic Med Sci, Nanchang, Jiangxi, Peoples R China 3.Univ Sci & Technol China, Hefei, Peoples R China 4.Anhui Univ, Inst Phys Sci & Informat Technol, Hefei, Peoples R China
推荐引用方式 GB/T 7714	Zhu, Lina,Zhao, Hongxin,Wang, Yizhuo,et al. Solubilization, purification, and ligand binding characterization of G protein-coupled receptor SMO in native membrane bilayer using styrene maleic acid copolymer[J]. PEERJ,2022,10.
APA	Zhu, Lina.,Zhao, Hongxin.,Wang, Yizhuo.,Yu, Chuandi.,Liu, Juanjuan.,...&Zhu, Lei.(2022).Solubilization, purification, and ligand binding characterization of G protein-coupled receptor SMO in native membrane bilayer using styrene maleic acid copolymer.PEERJ,10.
MLA	Zhu, Lina,et al."Solubilization, purification, and ligand binding characterization of G protein-coupled receptor SMO in native membrane bilayer using styrene maleic acid copolymer".PEERJ 10(2022).