Positive Effects of Isopropanol as a Co-Precipitant in Glycerol-3-Phosphate Acyltransferase Crystallization
文献类型：期刊论文
| 作者 | Liu, Yinghui1; Xue, Song1; Zhang, Yunxiu1,2; Feng, Yanbin1; Wang, Yayue1,2; Cao, Xupeng1 |
| 刊名 | JOURNAL OF OCEAN UNIVERSITY OF CHINA
 |
| 出版日期 | 2019-02-01 |
| 卷号 | 18期号:1页码:227-231 |
| ISSN号 | 1672-5182 |
| 关键词 | isopropanol polyethylene glycol (PEG) optimization glycerol-3-phosphate acyltransferase (GPAT) crystallization |
| DOI | 10.1007/s11802-019-3519-0 |
| 通讯作者 | Xue, Song(xuesong@dicp.ac.cn) |
| 英文摘要 | Glycerol-3-phosphate acyltransferase (GPAT) is considered as the rate-limiting enzyme of glycerolipid synthesis pathway and the core element in lysophosphatidic acid (LPA) synthesis. For understanding its catalytic mechanism, the structural biology study is expected, but is always hindered by obtaining crystals for X-ray diffraction analysis. In this study, a progressive strategy to optimize the crystal of microalgae plastidial GPAT was presented. After the expression and purification of GPAT, the crystals were screened by hanging-drop and only clusters were obtained. The crystals were optimized by adjusting temperature, pH, protein concentration, or precipitant, but little improvement. To improve the interaction between protein and precipitant, the isopropanol was applied as co-precipitant. The qualified crystals formed. It's suggested that isopropanol is critical to affect protein crystallization by altering polyethylene glycol (PEG)-water-protein interaction when PEG serves as precipitant. The resulting crystal diffracted to a resolution of 2.75 angstrom and belonged to space group P1, with unit-cell parameters a = 50.79, b = 80.09, c = 88.21 angstrom, and = 62.85, = 73.04, = 80.53 degrees. This work introduced a new strategy to optimize the crystal of the heterogeneous catalysis enzymes like GPAT and provided the fundamental structural information for the oriented synthesis of marine microalgae glycerolipid. |
| WOS关键词 | SN-GLYCEROL-3-PHOSPHATE ACYLTRANSFERASE ; EXPRESSION ; PROTEINS ; CLONING ; FAMILY ; GENE |
| 资助项目 | National Natural Science Foundation of China[21576253] ; National Natural Science Foundation of China[31500294] ; National Natural Science Foundation of China[31470432] |
| WOS研究方向 | Oceanography |
| 语种 | 英语 |
| 出版者 | OCEAN UNIV CHINA |
| WOS记录号 | WOS:000455483900023 |
| 资助机构 | National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; National Natural Science Foundation of China |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/166194]  |
| 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 | Xue, Song |
| 作者单位 | 1.Chinese Acad Sci, Dalian Inst Chem Phys, Marine Bioengn Grp, Dalian 116023, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Liu, Yinghui,Xue, Song,Zhang, Yunxiu,et al. Positive Effects of Isopropanol as a Co-Precipitant in Glycerol-3-Phosphate Acyltransferase Crystallization[J]. JOURNAL OF OCEAN UNIVERSITY OF CHINA,2019,18(1):227-231. |
| APA | Liu, Yinghui,Xue, Song,Zhang, Yunxiu,Feng, Yanbin,Wang, Yayue,&Cao, Xupeng.(2019).Positive Effects of Isopropanol as a Co-Precipitant in Glycerol-3-Phosphate Acyltransferase Crystallization.JOURNAL OF OCEAN UNIVERSITY OF CHINA,18(1),227-231. |
| MLA | Liu, Yinghui,et al."Positive Effects of Isopropanol as a Co-Precipitant in Glycerol-3-Phosphate Acyltransferase Crystallization".JOURNAL OF OCEAN UNIVERSITY OF CHINA 18.1(2019):227-231. |
入库方式： OAI收割
来源：大连化学物理研究所
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