Detection of metabolite-protein interactions in complex biological samples by high-resolution relaxometry: toward interactomics by NMR

Ziqing Wang (1) , Simone Pisano (1) , Veronica Ghini (2) , Pavel Kadeřávek (1) , Milan Zachrdla (1) , Philippe Pelupessy (3) , Morgan Kazmierczak (4) , Thorsten Marquardsen (5) , Jean-Max Tyburn (5) , Guillaume Bouvignies (1) , Giacomo Parigi (6) , Claudio Luchinat (6) , Fabien Ferrage (1)

1 LBM UMR 7203 - Laboratoire des biomolécules
2 CIRMMP - Consorzio Interuniversitario Risonanze Magnetiche di Metallo Proteine
3 LBM-E3 - Structure et Dynamique des Biomolécules
4 EPFL - Ecole Polytechnique Fédérale de Lausanne
5 Bruker BioSpin MRI GmbH [Ettlingen, Germany]
6 UniFI - Università degli Studi di Firenze = University of Florence = Université de Florence

Metabolomics, the systematic investigation of metabolites in biological fluids, cells or tissues, reveals essential information about metabolism and diseases. Metabolites have functional roles in myriads 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 hitherto still scarce. 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.

Mots clés: Metabolomics; Interactomics; NMR; Relaxometry

Domaines: Chimie

Ziqing Wang, Simone Pisano, Veronica Ghini, Pavel Kadeřávek, Milan Zachrdla, et al.. Detection of metabolite-protein interactions in complex biological samples by high-resolution relaxometry: toward interactomics by NMR. Journal of the American Chemical Society, 2021, 143 (25), pp.9393-9404. ⟨10.1021/jacs.1c01388⟩. ⟨hal-03291374⟩