Tailored nanodisc immobilization for one-step purification and reconstitution of cytochrome P450: A tool for membrane proteins' hard cases
文献类型：期刊论文
| 作者 | Zhao, Lan4; Tao, Jiaoli3,4; Huang, Yongdong4; Zhu, Kai4; Du, Yuxiang3,4; Hao, Dongxia4; Liu, Hongying3; Zhang, Rongyue2; Ma, Guanghui1,4 |
| 刊名 | JOURNAL OF SEPARATION SCIENCE
 |
| 出版日期 | 2021-08-06 |
| 页码 | 12 |
| 关键词 | cytochrome P450 membrane protein purification nanodiscs nanodisc-immobilized chromatography reconstitution |
| ISSN号 | 1615-9306 |
| DOI | 10.1002/jssc.202100284 |
| 英文摘要 | A novel nanodisc-based immobilization method was developed for high-efficient purification and reconstitution of cytochrome P450 in one step. Using membrane scaffold protein containing a histidine tag, charged-nanodiscs were prepared in the form of self-assembly of lipid-protein nanoparticles. Their properties including the particle diameter and its distribution and Zeta potential were controlled well by adjusting molar ratios of phospholipids to membrane scaffold protein. At an optimum lipid-to-membrane scaffold protein molar ratio of 60:1, uniformly regular-shaped and discoidal nanodiscs with an average particle diameter of 10 nm and Zeta potential of -19 mV were obtained. They can be well fractionated by size exclusion chromatography. Charged-nanodiscs were successfully immobilized onto Ni-chelating microspheres via histidine tags with a density of 6.6 mg membrane scaffold protein/mL gel. After being packed in a column, chromatography studies demonstrated that this nanodisc-immobilized chromatographic medium had a specific binding to cytochrome P450 in rat liver microsome. Nanodiscs containing cytochrome P450 can be furthermore eluted from the column with a diameter of about 87.0 nm and height of about 8.0 nm, respectively. The purity of cytochrome P450 after purification increased 25 folds strikingly. This nanodisc-immobilized chromatography method is promising for the one-step purification and reconstitution of membrane protein. |
| WOS关键词 | KINETIC-ANALYSIS ; BINDING ; PERFORMANCE ; CHROMATOGRAPHY ; EXTRACTION ; EXPRESSION ; PARTICLES ; MEDIA ; 3A4 |
| 资助项目 | National Key Research and Development Program of China[2018YFC0311101] ; Beijing Natural Science Foundation[L192017] ; National Natural Science Foundation of China[21821005] ; National Natural Science Foundation of China[22078334] ; National Natural Science Foundation of China[21902160] ; Youth Top Talent Project of Organization Department of Beijing Municipal Committee[ZZB2019001] |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:000681596600001 |
| 出版者 | WILEY-V C H VERLAG GMBH |
| 资助机构 | National Key Research and Development Program of China ; Beijing Natural Science Foundation ; National Natural Science Foundation of China ; Youth Top Talent Project of Organization Department of Beijing Municipal Committee |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/49596]  |
| 专题 | 中国科学院过程工程研究所 |
| 通讯作者 | Huang, Yongdong; Ma, Guanghui |
| 作者单位 | 1.Univ Chinese Acad Sci, Sch Chem Engn, Beijing, Peoples R China 2.Beijing Inst Petrochem Technol, Dept Appl Chem, Beijing, Peoples R China 3.China Univ Min & Technol, Sch Chem & Environm Engn, Beijing, Peoples R China 4.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhao, Lan,Tao, Jiaoli,Huang, Yongdong,et al. Tailored nanodisc immobilization for one-step purification and reconstitution of cytochrome P450: A tool for membrane proteins' hard cases[J]. JOURNAL OF SEPARATION SCIENCE,2021:12. |
| APA | Zhao, Lan.,Tao, Jiaoli.,Huang, Yongdong.,Zhu, Kai.,Du, Yuxiang.,...&Ma, Guanghui.(2021).Tailored nanodisc immobilization for one-step purification and reconstitution of cytochrome P450: A tool for membrane proteins' hard cases.JOURNAL OF SEPARATION SCIENCE,12. |
| MLA | Zhao, Lan,et al."Tailored nanodisc immobilization for one-step purification and reconstitution of cytochrome P450: A tool for membrane proteins' hard cases".JOURNAL OF SEPARATION SCIENCE (2021):12. |
入库方式： OAI收割
来源：过程工程研究所
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