Protein–Ligand Binding Volume Determined from a Single 2D NMR Spectrum with Increasing Pressure

• DOI: 10.1021/acs.jpcb.1c02917
• Publication Date: 2021-05-25T17:02:50Z
• Authors: Gediminas Skvarna...; Zigmantas Toleikis; Vilma Michailovienė; Christian Roumestand; Daumantas Matulis; Vytautas Petrauskas
• Keywords: Heteronuclear molecule; Protein ligand

Proteins undergo changes in their partial volumes numerous biological processes such as enzymatic catalysis, unfolding–refolding, and ligand binding. The change the protein volume upon binding—a parameter termed protein–ligand binding volume—can be extensively studied by high-pressure NMR spectroscopy. In this study, we developed a method to determine from single two-dimensional (2D) 1H–15N heteronuclear quantum coherence (HSQC) spectrum at different pressures, if exchange between ligand-free ligand-bound states of is slow time-scale. This approach required significantly lower amount time two carbonic anhydrase isozymes ligands. proposed can used other systems expand knowledge about small-molecule