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固定化酶的核磁共振(NMR)研究。

NMR of Immobilized Enzymes.

机构信息

Magnetic Resonance Center (CERM), University of Florence and Consorzio Interuniversitario, Risonanze Magnetiche di Metallo Proteine (CIRMMP), Sesto Fiorentino, Italy.

Department of Chemistry, University of Florence, Sesto Fiorentino, Italy.

出版信息

Methods Mol Biol. 2020;2100:363-383. doi: 10.1007/978-1-0716-0215-7_24.

Abstract

Solid-state NMR has become the method of choice for the assessment of protein structure for insoluble objects lacking long-range order. In this context, it is apparent that solid-state NMR is also perfectly poised toward the characterization of immobilized proteins. For these systems, it is possible to understand at the atomic level which perturbations, if any, are occurring as a result of the functionalization. Here we describe how it is possible to accomplish the NMR characterization of enzymes that have been immobilized through different approaches, and we introduce the reader to the choice of the experimental strategy that can be useful in different cases. An outlook on the level of information that can be attained is also given, in view of recent methodological advancements.

摘要

固态 NMR 已成为评估缺乏长程有序的不溶性物体中蛋白质结构的首选方法。在这种情况下,显然固态 NMR 也非常适合表征固定化蛋白质。对于这些系统,可以在原子水平上理解由于功能化而发生的任何扰动。在这里,我们描述了如何完成通过不同方法固定化的酶的 NMR 表征,并向读者介绍了在不同情况下可能有用的实验策略选择。鉴于最近的方法学进展,还对可以获得的信息量进行了展望。

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