Crystal Structures of Streptococcus suis Mannonate Dehydratase (ManD) and Its Complex with Substrate: Genetic and Biochemical Evidence for a Catalytic Mechanism
文献类型：期刊论文
| 作者 | Zhang, Qiangmin ; Gao, Feng ; Peng, Hao ; Cheng, Hao ; Liu, Yiwei ; Tang, Jiaqi ; Thompson, John ; Wei, Guohua ; Zhang, Jingren ; Du, Yuguo ; Yan, Jinghua ; Gao, George F. |
| 刊名 | JOURNAL OF BACTERIOLOGY
 |
| 出版日期 | 2009-09-15 |
| 卷号 | 191期号:18页码:5832-5837 |
| 中文摘要 | Mannonate dehydratase (ManD) is found only in certain bacterial species, where it participates in the dissimilation of glucuronate. ManD catalyzes the dehydration of D-mannonate to yield 2-keto-3-deoxygluconate (2-KDG), the carbon and energy source for growth. Selective inactivation of ManD by drug targeting is of therapeutic interest in the treatment of human Streptococcus suis infections. Here, we report the overexpression, purification, functional characterization, and crystallographic structure of ManD from S. suis. Importantly, by Fourier transform mass spectrometry, we show that 2-KDG is formed when the chemically synthesized substrate (D-mannonate) is incubated with ManD. Inductively coupled plasma-mass spectrometry revealed the presence of Mn(2+) in the purified protein, and in the solution state catalytically active ManD exists as a homodimer of two 41-kDa subunits. The crystal structures of S. suis ManD in native form and in complex with its substrate and Mn(2+) ion have been solved at a resolution of 2.9 angstrom. The core structure of S. suis ManD is a TIM barrel similar to that of other members of the xylose isomerase-like superfamily. Structural analyses and comparative amino acid sequence alignments provide evidence for the importance of His311 and Tyr325 in ManD activity. The results of site-directed mutagenesis confirmed the functional role(s) of these residues in the dehydration reaction and a plausible mechanism for the ManD-catalyzed reaction is proposed. |
| WOS记录号 | WOS:000269372600030 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/21477]  |
| 专题 | 生态环境研究中心_环境化学与生态毒理学国家重点实验室 |
| 推荐引用方式 GB/T 7714 | Zhang, Qiangmin,Gao, Feng,Peng, Hao,et al. Crystal Structures of Streptococcus suis Mannonate Dehydratase (ManD) and Its Complex with Substrate: Genetic and Biochemical Evidence for a Catalytic Mechanism[J]. JOURNAL OF BACTERIOLOGY,2009,191(18):5832-5837. |
| APA | Zhang, Qiangmin.,Gao, Feng.,Peng, Hao.,Cheng, Hao.,Liu, Yiwei.,...&Gao, George F..(2009).Crystal Structures of Streptococcus suis Mannonate Dehydratase (ManD) and Its Complex with Substrate: Genetic and Biochemical Evidence for a Catalytic Mechanism.JOURNAL OF BACTERIOLOGY,191(18),5832-5837. |
| MLA | Zhang, Qiangmin,et al."Crystal Structures of Streptococcus suis Mannonate Dehydratase (ManD) and Its Complex with Substrate: Genetic and Biochemical Evidence for a Catalytic Mechanism".JOURNAL OF BACTERIOLOGY 191.18(2009):5832-5837. |
入库方式： OAI收割
来源：生态环境研究中心
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。