Isotopic labelings for mechanistic studies.

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology
Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-02-07 DOI:10.1016/bs.mie.2024.01.011
Houchao Xu, Jeroen S Dickschat
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引用次数: 0

Abstract

The intricate mechanisms in the biosynthesis of terpenes belong to the most challenging problems in natural product chemistry. Methods to address these problems include the structure-based site-directed mutagenesis of terpene synthases, computational approaches, and isotopic labeling experiments. The latter approach has a long tradition in biosynthesis studies and has recently experienced a revival, after genome sequencing enabled rapid access to biosynthetic genes and enzymes. Today, this allows for a combined approach in which isotopically labeled substrates can be incubated with recombinant terpene synthases. These clearly defined reaction setups can give detailed mechanistic insights into the reactions catalyzed by terpene synthases, and recent developments have substantially deepened our understanding of terpene biosynthesis. This chapter will discuss the state of the art and introduce some of the most important methods that make use of isotopic labelings in mechanistic studies on terpene synthases.

用于机理研究的同位素标记。
萜烯的生物合成机制错综复杂,是天然产物化学领域最具挑战性的问题。解决这些问题的方法包括基于结构的萜烯合成酶定点诱变、计算方法和同位素标记实验。后一种方法在生物合成研究中有着悠久的传统,最近在基因组测序能够快速获取生物合成基因和酶之后又经历了复兴。如今,这种方法可以将同位素标记的底物与重组萜烯合成酶结合起来。这些定义明确的反应设置可以让我们从机理上详细了解萜烯合成酶催化的反应,最近的研究进展大大加深了我们对萜烯生物合成的了解。本章将讨论相关技术的发展状况,并介绍在萜烯合成酶机理研究中使用同位素标记的一些最重要的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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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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