Crystal structure of Bacillus fastidious uricase reveals an unexpected folding of the C-terminus residues crucial for thermostability under physiological conditions
文献类型：期刊论文
| 作者 | Feng, Juan4; Wang, Lu; Liu, Hongbo; Yang, Xiaolan; Liu, Lin4; Xie, Yanling; Liu, Miaomiao; Zhao, Yunsheng; Li, Xiang; Wang, Deqiang |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 2015 |
| 卷号 | 99期号:19页码:7973-7986 |
| 关键词 | Bacillus fastidious uricase C-terminus residues Thermal stability Electrostatic interaction center Hydrophobic interaction |
| ISSN号 | 0175-7598 |
| DOI | 10.1007/s00253-015-6520-6 |
| 文献子类 | Article |
| 英文摘要 | Bacillus fastidious uricase (BF uricase) containing 322 amino acid residues exhibited high stability under physiological conditions. Its crystal structure was solved to 1.4- resolution, showing homotetramer containing two homodimers. After the intersubunit antiparallel beta-sheet in its homodimer, each subunit had a total of 18 C-terminus residues forming an alpha-helix (Q305-A313) and random coil (S314-L322) on surface to bury other two alpha-helices (I227-T238 and I244-R258). In comparison, reported crystal structures of Arthrobacter globiformis and Aspergillus flavus uricases had atomic coordinates of only some C-terminus residues, while the crystal structures of all the other uricases accessible before September 2014 missed atomic coordinates of all their C-terminus residues, after the intersubunit antiparallel beta-sheets. In each homodimer of BF uricase, H-bonds were found between E311 and Y249 and between Y319 and D257; electrostatic interaction networks were found to surround D307 plus R310 and intersubunit R3, K312 plus D257, E318 plus K242, and L322 plus R258. Amino acid mutations that disrupted those interactions when R3 and D307 were reserved caused moderate decreases of activity at pH 9.2 while negligible decreases of activity at pH 7.4, but destroyed stability at pH 7.4 while slightly decreased stability at pH 9.2. Such structural information guided the fusion of 6His-tag to the C-terminus of the mutant L322D with SNSNSN as a linker to reserve the activity and stability. Hence, the folding of the C-terminus residues is crucial for thermal stability of BF uricase under physiological conditions; these new structural insights are valuable for molecular engineering of uricases. |
| 学科主题 | Biotechnology & Applied Microbiology |
| 出版地 | NEW YORK |
| 电子版国际标准刊号 | 1432-0614 |
| WOS关键词 | URATE OXIDASE ; ASPERGILLUS-FLAVUS ; ARTHROBACTER-GLOBIFORMIS ; POLY(ETHYLENE GLYCOL) ; RESOLUTION STRUCTURES ; STRUCTURE VALIDATION ; DIRECTED EVOLUTION ; MOLPROBITY ; EXPRESSION ; STABILITY |
| WOS研究方向 | Science Citation Index Expanded (SCI-EXPANDED) |
| 语种 | 英语 |
| WOS记录号 | WOS:000360839800015 |
| 出版者 | SPRINGER |
| 资助机构 | National Natural Science Foundation of China [30672139] ; Natural Sciences Foundation of CQ [CSTC2011BA5039, CSTC2012JJA0057] ; Education Ministry of China [20125503110007] ; National Science Foundation of USA [CHE-1111761] |
| 源URL | [http://ir.ibcas.ac.cn/handle/2S10CLM1/25893]  |
| 专题 | 中科院光生物学重点实验室 |
| 作者单位 | 1.Univ Kentucky, Coll Pharm, Mol Modeling & Biopharmaceut Ctr, Lexington, KY 40536 USA 2.Univ Kentucky, Coll Pharm, Dept Pharmaceut Sci, Lexington, KY 40536 USA 3.Chinese Acad Sci, Photosynth Res Ctr, Inst Bot, Key Lab Photobiol, Beijing 100093, Peoples R China 4.Chongqing Med Univ, Educ Minist, Coll Lab Med, Key Lab Med Lab Diagnost, Chongqing 400016, Peoples R China |
| 推荐引用方式 GB/T 7714 | Feng, Juan,Wang, Lu,Liu, Hongbo,et al. Crystal structure of Bacillus fastidious uricase reveals an unexpected folding of the C-terminus residues crucial for thermostability under physiological conditions[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2015,99(19):7973-7986. |
| APA | Feng, Juan.,Wang, Lu.,Liu, Hongbo.,Yang, Xiaolan.,Liu, Lin.,...&Liao, Fei.(2015).Crystal structure of Bacillus fastidious uricase reveals an unexpected folding of the C-terminus residues crucial for thermostability under physiological conditions.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,99(19),7973-7986. |
| MLA | Feng, Juan,et al."Crystal structure of Bacillus fastidious uricase reveals an unexpected folding of the C-terminus residues crucial for thermostability under physiological conditions".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 99.19(2015):7973-7986. |
入库方式： OAI收割
来源：植物研究所
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