Solubilization, purification, and ligand binding characterization of G protein-coupled receptor SMO in native membrane bilayer using styrene maleic acid copolymer
文献类型：期刊论文
| 作者 | Zhu, Lina3,4; Zhao, Hongxin3 ; Wang, Yizhuo1,3; Yu, Chuandi1,3; Liu, Juanjuan3; Li, Ling1,3; Li, Zehua1,3; Zhang, Jin2; Dai, Han3; Wang, Junfeng1,3,4
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| 刊名 | PEERJ
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| 出版日期 | 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. |
| WOS关键词 | MONOMERIC INTERLEUKIN-8 ; NANOPARTICLES ; NANODISCS ; RECONSTITUTION ; SIZE ; CHANNEL ; COMPLEX ; SMALPS ; CXCR1 |
| 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研究方向 | Science & Technology - Other Topics |
| 语种 | 英语 |
| WOS记录号 | WOS:000797871100004 |
| 出版者 | PEERJ INC |
| 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 |
| 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.Univ Sci & Technol China, Hefei, Peoples R China 2.Nanchang Univ, Sch Basic Med Sci, Nanchang, Jiangxi, Peoples R China 3.Chinese Acad Sci, Hefei Inst Phys Sci, High Magnet Field Lab, Key Lab High Magnet Field & Ion Beam Phys Biol, 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). |
入库方式： OAI收割
来源：合肥物质科学研究院
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