Structural characterization of the Pseudomonas aeruginosa dehydrogenase AtuB involved in citronellol and geraniol catabolism
Structural characterization of the Pseudomonas aeruginosa dehydrogenase AtuB involved in citronellol and geraniol catabolism
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参与香茅醇和香叶醇分解代谢的铜绿假单胞菌脱氢酶 AtuB 的结构表征
DOI：
10.1016/j.bbrc.2020.01.052
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发表时间：
2020
影响因子：
3.1
通讯作者：
Bartlam Mark
中科院分区：
文献类型：
--
作者：
Chen Yujing;Jia Haizhu;Liang Yakun;Zhang Hao;Che Shiyou;Liu Ruihua;Zhang Qionglin;Bartlam Mark
铜绿假单胞菌（Pseudomonasaerodynosacan）代谢非环状单萜类化合物（如香茅醇和香叶醇）作为唯一的碳源和能源。在铜绿假单胞菌（Pseudomonasaerosaas）中共鉴定出7种蛋白（AtuA、AtuB、AtuCF、AtuD、AtuE、AtuG、AtuH）参与非环萜的利用途径。AtuB是一种脱氢酶，在无环萜烯利用（Atu）途径中负责香茅醇和香叶醇催化，尽管其结构和功能迄今尚未被表征。本文报道了铜绿假单胞菌PAO 1（PaAtuB）AtuB的晶体结构，其分辨率为1.8 nm。PaAtuB在空间群F222中结晶，在不对称单元中具有单个单体。分析超离心数据显示PaAtuB在溶液中形成稳定的四聚体，这与结构一致。结构分析证实AtuB属于短链脱氢酶/还原酶（SDR）家族。根据晶体结构预测AtuB与NADP（H）结合，这一点通过MicroScale热泳分析得到证实，该分析显示PaAtuB与NADP（H）结合的Kd值为258 μM。这项工作为探索AtuB的潜在生物技术和制药应用提供了一个起点。
Pseudomonas aeruginosacan metabolize acyclic monoterpenoids (such as citronellol and geraniol) as the only carbon and energy sources. A total of seven proteins (AtuA, AtuB, AtuCF, AtuD, AtuE, AtuG, AtuH) have been identified inPseudomonas aeruginosaas participating in the acyclic terpene utilization pathway. AtuB is a dehydrogenase enzyme responsible for citronellol and geraniol catabolism in the acyclic terpene utilization (Atu) pathway, although its structure and function have not been characterized to date. Here we report the crystal structure of AtuB fromPseudomonas aeruginosaPAO1 (PaAtuB) to 1.8 Å resolution. PaAtuB crystallizes in the space group F222 with a single monomer in the asymmetric unit. Analytical ultracentrifugation data shows that PaAtuB forms a stable tetramer in solution, which is consistent with the structure. Structural analysis confirms that AtuB belongs to the short-chain dehydrogenase/reductase (SDR) family. AtuB is predicted to bind NADP(H) from the crystal structure, which is confirmed by MicroScale Thermophoresis analysis that shows PaAtuB binds NADP(H) with a Kd value of 258 μM. This work provides a starting point to explore potential biotechnology and pharmaceutical applications of AtuB.