Structure of the ent-Copalyl Diphosphate Synthase PtmT2 from Streptomyces platensis CB00739, a Bacterial Type II Diterpene Synthase.
Structure of the ent-Copalyl Diphosphate Synthase PtmT2 from Streptomyces platensis CB00739, a Bacterial Type II Diterpene Synthase.
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DOI：
10.1021/jacs.6b04317
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发表时间：
2016-08-31
影响因子：
15
通讯作者：
Shen B
中科院分区：
文献类型：
--
作者：
Rudolf JD;Dong LB;Cao H;Hatzos-Skintges C;Osipiuk J;Endres M;Chang CY;Ma M;Babnigg G;Joachimiak A;Phillips GN Jr;Shen B
萜类化合物是自然界中发现的最大和结构最多样化的天然产物家族，但它们在细菌中的存在却被低估了。萜类化合物的碳骨架是通过萜类糖苷酶（TS）催化的碳阳离子依赖性环化级联反应产生的。I型和II型TS分别通过二磷酸离子化和质子化引发环化，并且两种类型的蛋白质结构都是已知的。大多数植物二萜脱氢酶（DTSs）具有三个α-螺旋结构域（αβγ），这被认为是由离散的祖先细菌I型TS（α）和II型TS（βγ）融合而产生的。细菌来源的II型DTS，其中没有结构特征的成员，是TS结构进化中缺失的部分。在这里，我们报告的第一个晶体结构的II型细菌。PtmT 2来源于钝顶链霉菌CB 00739，是一种对映柯巴基二磷酸合成酶，参与了Platensiomycin和Platencin的生物合成。PtmT 2的晶体结构在1.80 nm的分辨率下得到解决，对接研究表明geranylgeranyl diphosphate（GGPP）的催化活性构象。定点诱变证实了参与结合GGPP的二磷酸部分的残基，并确定DxxxxE作为细菌来源的II型DTS的潜在Mg 2+结合基序。最后，形状和物理化学性质的活性位点是负责确定特定的催化结果的TS。PtmT 2的结构从根本上推进了细菌TS，其机制及其在TS进化中的作用的知识。
Terpenoids are the largest and most structurally diverse family of natural products found in nature, yet their presence in bacteria is underappreciated. The carbon skeletons of terpenoids are generated through carbocation-dependent cyclization cascades catalyzed by terpene synthases (TSs). Type I and type II TSs initiate cyclization via diphosphate ionization and protonation, respectively, and protein structures of both types are known. Most plant diterpene synthases (DTSs) possess three α-helical domains (αβγ), which are thought to have arisen from the fusion of discrete, ancestral bacterial type I TSs (α) and type II TSs (βγ). Type II DTSs of bacterial origin, of which there are no structurally characterized members, are a missing piece in the structural evolution of TSs. Here, we report the first crystal structure of a type II DTS from bacteria. PtmT2 from Streptomyces platensis CB00739 was verified as an ent-copalyl diphosphate synthase involved in the biosynthesis of platensimycin and platencin. The crystal structure of PtmT2 was solved at a resolution of 1.80 Å, and docking studies suggest the catalytically active conformation of geranylgeranyl diphosphate (GGPP). Site-directed mutagenesis confirmed residues involved in binding the diphosphate moiety of GGPP and identified DxxxxE as a potential Mg2+-binding motif for type II DTSs of bacterial origin. Finally, both the shape and physicochemical properties of the active sites are responsible for determining specific catalytic outcomes of TSs. The structure of PtmT2 fundamentally advances the knowledge of bacterial TSs, their mechanisms, and their role in the evolution of TSs.
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影响因子：
4.8
作者：
Kim, Youngchang;Babnigg, Gyorgy;Jedrzejczak, Robert;Eschenfeldt, William H.;Li, Hui;Maltseva, Natalia;Hatzos-Skintges, Catherine;Gu, Minyi;Makowska-Grzyska, Magdalena;Wu, Ruiying;An, Hao;Chhor, Gekleng;Joachimiak, Andrzej
通讯作者：
Joachimiak, Andrzej
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
影响因子：
7.4
作者：
Keeling, Christopher I.;Dullat, Harpreet K.;Bohlmann, Joerg
通讯作者：
Bohlmann, Joerg