Wang Ziqing, Pisano Simone, Ghini Veronica, Kadeřávek Pavel, Zachrdla Milan, Pelupessy Philippe, Kazmierczak Morgan, Marquardsen Thorsten, Tyburn Jean-Max, Bouvignies Guillaume, Parigi Giacomo, Luchinat Claudio, Ferrage Fabien
Laboratoire des Biomolécules, LBM, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, CNRS, 75005 Paris, France.
Consorzio Interuniversitario Risonanze Magnetiche Metallo Proteine (CIRMMP), via Sacconi 6, Sesto Fiorentino, 50019 Italy.
J Am Chem Soc. 2021 Jun 16. doi: 10.1021/jacs.1c01388.
Metabolomics, the systematic investigation of metabolites in biological fluids, cells, or tissues, reveals essential information about metabolism and diseases. Metabolites have functional roles in a myriad of biological processes, as substrates and products of enzymatic reactions but also as cofactors and regulators of large numbers of biochemical mechanisms. These functions involve interactions of metabolites with macromolecules. Yet, methods to systematically investigate these interactions are still scarce to date. In particular, there is a need for techniques suited to identify and characterize weak metabolite-macromolecule interactions directly in complex media such as biological fluids. Here, we introduce a method to investigate weak interactions between metabolites and macromolecules in biological fluids. Our approach is based on high-resolution NMR relaxometry and does not require any invasive procedure or separation step. We show that we can detect interactions between small and large molecules in human blood serum and quantify the size of the complex. Our work opens the way for investigations of metabolite (or other small molecules)-protein interactions in biological fluids for interactomics or pharmaceutical applications.
代谢组学是对生物体液、细胞或组织中的代谢物进行系统研究，它揭示了有关代谢和疾病的重要信息。代谢物在众多生物过程中发挥着功能性作用，既是酶促反应的底物和产物，也是大量生化机制的辅助因子和调节剂。这些功能涉及代谢物与大分子的相互作用。然而，迄今为止，系统研究这些相互作用的方法仍然很少。特别是，需要适合在诸如生物体液等复杂介质中直接鉴定和表征弱代谢物 - 大分子相互作用的技术。在此，我们介绍一种研究生物体液中代谢物与大分子之间弱相互作用的方法。我们的方法基于高分辨率核磁共振弛豫测量法，不需要任何侵入性程序或分离步骤。我们表明，我们可以检测人血清中小分子与大分子之间的相互作用，并量化复合物的大小。我们的工作为在生物体液中进行代谢物（或其他小分子） - 蛋白质相互作用的研究开辟了道路，可用于相互作用组学或药物应用。
