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

• Published in: The journal of physical chemistry. B Vol. 125; no. 22; pp. 5823 - 5831
• Main Authors: Skvarnavičius, Gediminas, Toleikis, Zigmantas, Michailovienė, Vilma, Roumestand, Christian, Matulis, Daumantas, Petrauskas, Vytautas
• Format: Journal Article
• Language: English
• Published: American Chemical Society 10.06.2021
• Subjects: B: Biophysical and Biochemical Systems and Processes; Life Sciences; Physics
• ISSN: 1520-6106; 1520-5207
• DOI: 10.1021/acs.jpcb.1c02917

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