Site-directed spin labeling-electron paramagnetic resonance spectroscopy in biocatalysis: Enzyme orientation and dynamics in nanoscale confinement
Site-directed spin labeling-electron paramagnetic resonance spectroscopy in biocatalysis: Enzyme orientation and dynamics in nanoscale confinement
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DOI：
10.1016/j.checat.2021.03.005
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发表时间：
2021-06-17
期刊：
影响因子：
--
通讯作者：
Yang, Zhongyu
中科院分区：
文献类型：
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作者：
Pan, Yanxiong;Li, Hui;Yang, Zhongyu
定点自旋标记（SDSL）与电子顺磁共振（EPR）光谱学相结合，可以探测复杂生物系统中原本难以获得的结构信息。我们最近扩展了SDSL-EPR，以揭示酶在介孔纳米结构中封装后的相对取向和骨架动力学，这为所观察到的生物催化活性奠定了结构基础。我们的策略引起了生物催化界的兴趣，因此在这篇资源文章中，我们介绍了将SDSL-EPR推广到非均相生物催化的原理和实验程序。我们将重点介绍介孔材料中的酶，并举例说明潜在陷阱的方法和注意事项。最终目标是为生物催化界提供一个强有力的资源，以填补长期存在的知识空白，在非均相生物催化和酶的结构-功能关系的酶-介孔材料的界面，并利用结构的见解来指导多孔平台的酶固定化的合理设计。
Site-directed spin labeling (SDSL) in combination with electron paramagnetic resonance (EPR) spectroscopy probes the otherwise inaccessible structural information in complex biological systems. We recently extended SDSL-EPR to reveal the relative orientation and backbone dynamics of enzymes upon encapsulation in mesoporous nanostructures, which set the structural basis underlying the observed biocatalytic activity. Our strategy had generated interest in the biocatalysis community, and thus in this resource article, we contribute an introduction to the principles and experimental procedure that generalize SDSL-EPR to heterogeneous biocatalysis. We will focus on enzymes in mesoporous materials with examples demonstrating the methods and cautions of potential pitfalls. The ultimate goal is to provide the biocatalysis community with a powerful resource to fill in a long-standing knowledge gap in heterogeneous biocatalysis and the structure-function relationship of enzymes at the interface of enzyme-mesoporous materials and utilize the structural insights to guide the rational design of porous platforms for enzyme immobilization.