Imaging Saturation Transfer Difference (STD) NMR: affinity and specificity of protein-ligand interactions from a single NMR sample

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Monaco, Serena, Angulo, Jesus and Wallace, Matthew (2023) Imaging Saturation Transfer Difference (STD) NMR: affinity and specificity of protein-ligand interactions from a single NMR sample. Journal of the American Chemical Society, 145 (30). 16391–16397. ISSN 0002-7863

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Abstract

We have combined saturation transfer difference NMR (STD NMR) with chemical shift imaging (CSI) and controlled concentration gradients of small molecule ligands to develop imaging STD NMR, a new tool for the assessment of protein-ligand interactions. Our methodology allows the determination of protein-ligand dissociation constants (KD) and assessment of the binding specificity in a single NMR tube, avoiding time-consuming titrations. We demonstrate the formation of suitable and reproducible concentration gradients of ligand along the vertical axis of the tube, against homogeneous protein concentration, and present a CSI pulse sequence for the acquisition of STD NMR experiments at different positions along the sample tube. Compared to the conventional methodology in which the [ligand]/[protein] ratio is increased manually, we can perform STD NMR experiments at a greater number of ratios and construct binding epitopes in a fraction (∼20%) of the experimental time. Second, imaging STD NMR also allows us to screen for non-specific binders, by monitoring any variation of the binding epitope map at increasing [ligand]/[protein] ratios. Hence, the proposed method does carry the potential to speed up and smooth out the drug discovery process.

Item Type: Article
Uncontrolled Keywords: catalysis,chemistry(all),biochemistry,colloid and surface chemistry ,/dk/atira/pure/subjectarea/asjc/1500/1503
Faculty \ School: Faculty of Science > School of Pharmacy (former - to 2024)
UEA Research Groups: Faculty of Science > Research Groups > Pharmaceutical Materials and Soft Matter
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Depositing User: LivePure Connector
Date Deposited: 28 Jul 2023 08:56
Last Modified: 02 Nov 2025 05:31
URI: https://ueaeprints.uea.ac.uk/id/eprint/92735
DOI: 10.1021/jacs.3c02218

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