Solid-state NMR protocols for unveiling dynamics and (drug) interactions of membrane-bound proteins.

IF 4.5 3区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Protein Science Pub Date : 2025-04-01 DOI:10.1002/pro.70102
Alessia Lasorsa, Patrick C A van der Wel
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引用次数: 0

Abstract

Magic angle spinning solid-state NMR (MAS ssNMR) is a versatile tool for studying the structure and dynamics of membrane proteins, as well as their interactions with ligands and drugs. Its power lies in the ability to provide atomic-level information on samples under physiological-like conditions. Moreover, it can illuminate dynamics across a wide range of timescales with great relevance to membrane protein function and dysfunction. In this protocol paper, we highlight key aspects of sample preparation, data acquisition, and interpretation, based on our own experience and the broader literature. We discuss key protocol steps along with important considerations for sample preparation and parameters for ssNMR measurements, with reference to the special requirements of membrane-based samples. Such samples display physiologically relevant dynamics across different motional regimes that can be probed by NMR but also can interfere with certain NMR measurements. We guide the reader through the whole process from sample preparation to complex NMR characterization techniques. Throughout the report, we refer back to examples from our own prior work on the interactions between cytochrome c and cardiolipin-containing membranes, with a discussion of the lipid dependence and interactions with a peroxidase-activity inhibitor. We conclude with a short discussion of alternative and new methods that are further boosting the power and versatility of ssNMR as a tool to study membrane-bound proteins and their ligands or drug interactions.

揭示膜结合蛋白动力学和(药物)相互作用的固态核磁共振协议。
魔角旋转固体核磁共振(MAS ssNMR)是研究膜蛋白结构和动力学及其与配体和药物相互作用的多功能工具。它的强大之处在于能够在类似生理的条件下提供样品的原子级信息。此外,它可以阐明与膜蛋白功能和功能障碍密切相关的大范围时间尺度的动力学。在这份协议文件中,我们根据自己的经验和广泛的文献,强调了样品制备、数据采集和解释的关键方面。我们讨论了关键的协议步骤,以及样品制备和ssNMR测量参数的重要考虑因素,并参考了膜基样品的特殊要求。这些样品在不同的运动机制中显示出生理相关的动态,可以通过核磁共振探测,但也可以干扰某些核磁共振测量。我们引导读者通过整个过程从样品制备到复杂的核磁共振表征技术。在整个报告中,我们回顾了我们自己之前关于细胞色素c和含心磷脂膜之间相互作用的研究,并讨论了脂质依赖性和与过氧化物酶活性抑制剂的相互作用。最后,我们简要讨论了一些替代方法和新方法,这些方法进一步提高了ssmr作为研究膜结合蛋白及其配体或药物相互作用的工具的能力和通用性。
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
Protein Science
Protein Science 生物-生化与分子生物学
CiteScore
12.40
自引率
1.20%
发文量
246
审稿时长
1 months
期刊介绍: Protein Science, the flagship journal of The Protein Society, is a publication that focuses on advancing fundamental knowledge in the field of protein molecules. The journal welcomes original reports and review articles that contribute to our understanding of protein function, structure, folding, design, and evolution. Additionally, Protein Science encourages papers that explore the applications of protein science in various areas such as therapeutics, protein-based biomaterials, bionanotechnology, synthetic biology, and bioelectronics. The journal accepts manuscript submissions in any suitable format for review, with the requirement of converting the manuscript to journal-style format only upon acceptance for publication. Protein Science is indexed and abstracted in numerous databases, including the Agricultural & Environmental Science Database (ProQuest), Biological Science Database (ProQuest), CAS: Chemical Abstracts Service (ACS), Embase (Elsevier), Health & Medical Collection (ProQuest), Health Research Premium Collection (ProQuest), Materials Science & Engineering Database (ProQuest), MEDLINE/PubMed (NLM), Natural Science Collection (ProQuest), and SciTech Premium Collection (ProQuest).
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