Speeding-up the Determination of Protein-Ligand Affinities by STD NMR: The Reduced Dataset STD NMR Approach (rd-STD NMR)
IIQ
Gabriel Rocha, Jonathan Ramírez-Cárdenas, M. Carmen Padilla-Pérez, Samuel Walpole, Ridvan Nepravishta, M. Isabel García-Moreno, Elena M. Sánchez-Fernández, Carmen Ortiz Mellet, Jesús Angulo, Juan C. Muñoz-García Anal. Chem. 2024, 96, 615−619

STD NMR spectroscopy is a powerful ligand-observed NMR tool for screening and characterizing the interactions of small molecules and low molecular weight fragments with a given macromolecule, identifying the main intermolecular contacts in the bound state. It is also a powerful analytical technique for the accurate determination of protein–ligand dissociation constants (KD) of medium-to-weak affinity, of interest in the pharmaceutical industry. However, accurate KD determination and epitope mapping requires a long series of experiments at increasing saturation times to carry out a full analysis using the so-called STD NMR build-up curve approach and apply the “initial slopes approximation”. Here, we have developed a new protocol to bypass this important limitation, which allows us to obtain initial slopes by using just two saturation times and, hence, to very quickly determine precise protein–ligand dissociation constants by STD NMR.

https://pubs.acs.org/doi/10.1021/acs.analchem.3c03980

Gabriel Rocha Domínguez y Jonathan Ramírez Cárdenas

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