Structure-based design of a hyperthermostable AgUricase for hyperuricemia and gout therapy
文献类型：期刊论文
| 作者 | Shi, Yi2 ; Wang, Ting2,3,4; Zhou, X. Edward1; Liu, Qiu-Feng2; Jiang, Yi2,3; Xu, H. Eric1,2,3,4
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| 刊名 | ACTA PHARMACOLOGICA SINICA
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| 出版日期 | 2019-10-01 |
| 卷号 | 40期号:10页码:1364-1372 |
| 关键词 | uricase thermostability disulfide cross-linking crystal structure |
| ISSN号 | 1671-4083 |
| DOI | 10.1038/s41401-019-0269-x |
| 通讯作者 | Shi, Yi(shiyi@simm.ac.cn) ; Xu, H. Eric(eric.xu@simm.ac.cn) |
| 英文摘要 | Arthrobacter globiformis Uricase (AgUricase) is a homotetrameric uricase with the potential for therapeutic use in treating hyperuricemia-related diseases. To achieve sufficient therapeutic effects, it is essential for this enzyme to have high thermostability and long half-life in physiological condition. To improve the thermostability of this enzyme, we introduced a series of cysteine pair mutations into the AgUricase subunits based on its structural model and studied the thermostability of the mutant enzymes with introduced disulfide bridges. Two intersubunit cysteine pair mutations, K12C-E286C and S296C-S296C, were found to markedly increase the melting temperatures of the corresponding mutant enzymes compared with WT AgUricase. The crystal structure of the K12C-E286C mutant at 1.99 angstrom resolution confirmed the formation of a distinct disulfide bond between the two subunits in the dimer. Structural analysis and biochemical data revealed that the C-terminal loop of AgUricase was flexible, and its interaction with neighboring subunits was required for the stability of the enzyme. We introduced an additional intersubunit K244C-C302 disulfide bond based on the crystal structure of the K12C-E286C mutant and confirmed that this additional disulfide bond further stabilized the flexible C-terminal loop and improved the thermostability of the enzyme. Disulfide cross-linking also protected AgUricase from protease digestion. Our studies suggest that the introduction of disulfide bonds into proteins is a potential strategy for enhancing the thermostability of multimeric proteins for medical applications. |
| WOS关键词 | THERMAL-STABILITY ; URATE OXIDASE ; DISULFIDE BONDS ; IMMUNOGENICITY ; DENATURATION ; PEGLOTICASE ; RASBURICASE ; EFFICACY ; URICASE ; SYSTEMS |
| 资助项目 | National Natural Science Foundation of China[31770796] ; Strategic Priority Research Program of CAS[XDB08020303] ; National Science and Technology Major Project[2018ZX09711002-002-002] ; K.C. Wong Education Foundation |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| WOS记录号 | WOS:000488273100011 |
| 出版者 | NATURE PUBLISHING GROUP |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/282671]  |
| 专题 | 中国科学院上海药物研究所 |
| 通讯作者 | Shi, Yi; Xu, H. Eric |
| 作者单位 | 1.Van Andel Res Inst, Lab Struct Sci, Grand Rapids, MI 49503 USA 2.Chinese Acad Sci, CAS Key Lab Receptor Res, VARI SIMM Ctr, Ctr Struct & Funct Drug Targets,Shanghai Inst Mat, Shanghai 201203, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201213, Peoples R China |
| 推荐引用方式 GB/T 7714 | Shi, Yi,Wang, Ting,Zhou, X. Edward,et al. Structure-based design of a hyperthermostable AgUricase for hyperuricemia and gout therapy[J]. ACTA PHARMACOLOGICA SINICA,2019,40(10):1364-1372. |
| APA | Shi, Yi,Wang, Ting,Zhou, X. Edward,Liu, Qiu-Feng,Jiang, Yi,&Xu, H. Eric.(2019).Structure-based design of a hyperthermostable AgUricase for hyperuricemia and gout therapy.ACTA PHARMACOLOGICA SINICA,40(10),1364-1372. |
| MLA | Shi, Yi,et al."Structure-based design of a hyperthermostable AgUricase for hyperuricemia and gout therapy".ACTA PHARMACOLOGICA SINICA 40.10(2019):1364-1372. |
入库方式： OAI收割
来源：上海药物研究所
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