Structural biology of terpene synthases.

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-03-31 DOI:10.1016/bs.mie.2024.03.012
Baiying Xing, Zhenyu Lei, Zhaoye Bai, Guowei Zang, Yuxian Wang, Chenyu Zhang, Minren Chen, Yucheng Zhou, Jiahao Ding, Donghui Yang, Ming Ma
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Abstract

Structural biology research of terpene synthases (TSs) has provided a useful basis to understand their catalytic mechanisms in producing diverse terpene products with polycyclic ring systems and multiple chiral centers. However, compared to the large numbers of>95,000 terpenoids discovered to date, few structures of TSs have been solved and the understanding of their catalytic mechanisms is lagging. We here (i) introduce the basic catalytic logic, the structural architectures, and the metal-binding conserved motifs of TSs; (ii) provide detailed experimental procedures, in gene cloning and plasmid construction, protein purification, crystallization, X-ray diffraction data collection and structural elucidation, for structural biology research of TSs; and (iii) discuss the prospects of structure-based engineering and de novo design of TSs in generating valuable terpene molecules, which cannot be easily achieved by chemical synthesis.

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萜烯合成酶的结构生物学。
萜烯合成酶(TSs)的结构生物学研究为了解其催化机制提供了有用的基础,这些催化机制可产生具有多环环系统和多个手性中心的多种萜烯产品。然而,与迄今为止发现的超过 95,000 种萜类化合物相比,已解决的 TSs 结构很少,对其催化机理的了解也很滞后。在此,我们(i) 介绍了 TSs 的基本催化逻辑、结构体系和金属结合保守基团;(ii) 提供了 TSs 结构生物学研究的详细实验步骤,包括基因克隆和质粒构建、蛋白质纯化、结晶、X 射线衍射数据收集和结构阐释;(iii) 讨论了基于结构的 TSs 工程设计和从头设计的前景,以生成有价值的萜烯分子,而这是化学合成难以实现的。
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来源期刊
Methods in enzymology
Methods in enzymology 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
308
审稿时长
3-6 weeks
期刊介绍: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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