Structure-guided product determination of the bacterial type II diterpene synthase Tpn2.
Structure-guided product determination of the bacterial type II diterpene synthase Tpn2.
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DOI：
10.1038/s42004-022-00765-6
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发表时间：
2022-11-08
影响因子：
5.9
通讯作者：
Rudolf, Jeffrey D.
中科院分区：
文献类型：
--
作者：
Stowell, Emma A.;Ehrenberger, Michelle A.;Lin, Ya-Lin;Chang, Chin-Yuan;Rudolf, Jeffrey D.
萜烯合成酶(TS)酶学的一个重大挑战是从蛋白质序列预测功能的能力。鉴于特征性细菌TS的数量有限，而且它们与真核同行之间存在显著的序列差异，目前这是不可能的。为了有助于理解II型细菌TSS的序列-结构-功能关系，我们测定了北孢子菌Tpn2的结构。对CB02891进行了X射线结晶学分析，并通过结构导向突变体对其作用机制进行了初步探讨。将甘氨酸替换为碱性残基，将产物的偏好从十字烷骨架改变为齐聚拉布丹骨架，导致了第一个从细菌TS中鉴定出的齐聚拉布丹。了解单个残基如何决定Tpn2中的环化模式，以及对细菌II型TSS进行详细的生物信息学分析，为研究细菌II型TSS的功能范围和发现新的细菌萜类化合物奠定了基础。阐明萜类合酶的晶体结构对于理解该类酶的序列-结构-功能关系具有重要意义。在这里，作者确定了细菌萜类合成酶Tpn2的晶体结构，并通过结构导向突变探索了其催化机理，并将产物偏好从景天烷骨架改变为正丁烷骨架。
A grand challenge in terpene synthase (TS) enzymology is the ability to predict function from protein sequence. Given the limited number of characterized bacterial TSs and significant sequence diversities between them and their eukaryotic counterparts, this is currently impossible. To contribute towards understanding the sequence-structure-function relationships of type II bacterial TSs, we determined the structure of the terpentedienyl diphosphate synthase Tpn2 from Kitasatospora sp. CB02891 by X-ray crystallography and made structure-guided mutants to probe its mechanism. Substitution of a glycine into a basic residue changed the product preference from the clerodane skeleton to a syn-labdane skeleton, resulting in the first syn-labdane identified from a bacterial TS. Understanding how a single residue can dictate the cyclization pattern in Tpn2, along with detailed bioinformatics analysis of bacterial type II TSs, sets the stage for the investigation of the functional scope of bacterial type II TSs and the discovery of novel bacterial terpenoids. Crystal structure elucidation of terpene synthases plays an important role to understand the sequence-structure-function relationships of this enzyme class. Here, the authors determine the crystal structure of bacterial terpene synthase Tpn2, probe the catalytic mechanism by structure-guided mutation, and change the product preference from a clerodane skeleton to a syn-labdane skeleton.
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影响因子：
14.9
作者：
Goujon M;McWilliam H;Li W;Valentin F;Squizzato S;Paern J;Lopez R
通讯作者：
Lopez R
影响因子：
16.1
作者：
Chen, Xiaoying;Zaro, Jennica L.;Shen, Wei-Chiang
通讯作者：
Shen, Wei-Chiang
DOI：
10.1351/pac-con-09-09-37
发表时间：
2010
期刊：
Pure and applied chemistry. Chimie pure et appliquee
影响因子：
--
作者：
Aaron JA;Christianson DW
通讯作者：
Christianson DW
影响因子：
2.9
作者：
Cao, Rong;Zhang, Yonghui;Mann, Francis M.;Huang, Cancan;Mukkamala, Dushyant;Hudock, Michael P.;Mead, Matthew E.;Prisic, Sladjana;Wang, Ke;Lin, Fu-Yang;Chang, Ting-Kai;Peters, Reuben J.;Odfield, Eric
通讯作者：
Odfield, Eric
影响因子：
14.9
作者：
ASLANIDIS, C;DEJONG, PJ
通讯作者：
DEJONG, PJ