"restoration" of glutathione transferase activity by single-site mutation of the yeast prion protein ure2
文献类型：期刊论文
| 作者 | Zhang, Zai-Rong1,2; Bai, Ming1; Wang, Xin-Yu1,3; Zhou, Jun-Mei1; Perrett, Sarah1 |
| 刊名 | Journal of molecular biology
 |
| 出版日期 | 2008-12-19 |
| 卷号 | 384期号:3页码:641-651 |
| 关键词 | Amyloid Ure2p Glutathione s-transferase Protein engineering Enzyme |
| ISSN号 | 0022-2836 |
| DOI | 10.1016/j.jmb.2008.09.047 |
| 通讯作者 | Perrett, sarah(sarah.perrett@iname.com) |
| 英文摘要 | The yeast prion ure2p is composed of an n-terminal prion domain, and a c-terminal globular domain, which shows similarity to glutathione transferases (gsts) in both sequence and structure. ure2p protects saccharomyces cerevisiae cells from heavy metal ion and oxidant toxicity. ure2p shows glutathione-dependent peroxidase (gpx) activity, which is often an adjunct activity of gsts, but wild-type ure2p shows no detectable gst activity toward the standard substrate 1-chloro-2,4-dinitrobenzene (cdnb). the structural basis for the substrate specificity of ure2p enzymatic activity is an interesting problem that is fundamental to understanding the in vivo roles of ure2p and its relationship to the gst structural family. the critical catalytic residue in the other known gsts is ser, tyr or cys. here, we demonstrate that residue n124 is important for the gpx activity of ure2p, and a wildtype level of activity is maintained in n124s, but not in n124y/c. interestingly, we found that the single-site mutations a122c and n124a/v (but not n124s/y/c) "restore" the gst activity of ure2p toward cdnb, while causing a substantial reduction in gpx activity. the steady-state kinetics for the gst activity of a122c appears to follow a ping-pong mechanism. in contrast, the gst activity of 124-site mutants shows a sequential mechanism,. as is observed for the native gpx activity of ure2p, and typical gst enzymes. these findings shed light on the evolutionary relationship of ure2p with other gst family members, and contribute to our understanding of catalytic promiscuity and divergent evolution. (c) 2008 elsevier ltd. all rights reserved. |
| WOS关键词 | SACCHAROMYCES-CEREVISIAE ; S-TRANSFERASE ; CATALYTIC PROMISCUITY ; AMYLOID FORMATION ; EVOLUTION ; PEROXIDASE ; DOMAIN ; SUPERFAMILY ; THIOREDOXIN ; DIVERSITY |
| WOS研究方向 | Biochemistry & Molecular Biology |
| WOS类目 | Biochemistry & Molecular Biology |
| 语种 | 英语 |
| WOS记录号 | WOS:000261711300008 |
| 出版者 | ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2386976 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Perrett, Sarah |
| 作者单位 | 1.Chinese Acad Sci, Natl Lab Biomacromol, Inst Biophys, Beijing 100101, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.Nankai Univ, Dept Biophys, Inst Phys, Tianjin 300071, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Zai-Rong,Bai, Ming,Wang, Xin-Yu,et al. "restoration" of glutathione transferase activity by single-site mutation of the yeast prion protein ure2[J]. Journal of molecular biology,2008,384(3):641-651. |
| APA | Zhang, Zai-Rong,Bai, Ming,Wang, Xin-Yu,Zhou, Jun-Mei,&Perrett, Sarah.(2008)."restoration" of glutathione transferase activity by single-site mutation of the yeast prion protein ure2.Journal of molecular biology,384(3),641-651. |
| MLA | Zhang, Zai-Rong,et al.""restoration" of glutathione transferase activity by single-site mutation of the yeast prion protein ure2".Journal of molecular biology 384.3(2008):641-651. |
入库方式： iSwitch采集
来源：中国科学院大学
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。