Determination of intracellular protein-ligand binding affinity by competition binding in-cell NMR.

ABSTRACT: Structure-based drug development suffers from high attrition rates due to the poor activity of lead compounds in cellular and animal models caused by low cell penetrance, off-target binding or changes in the conformation of the target protein in the cellular environment. The latter two effects cause a change in the apparent binding affinity of the compound, which is indirectly assessed by cellular activity assays. To date, direct measurement of the intracellular binding affinity remains a challenging task. In this work, in-cell NMR spectroscopy was applied to measure intracellular dissociation constants in the nanomolar range by means of protein-observed competition binding experiments. Competition binding curves relative to a reference compound could be retrieved either from a series of i

SUBMITTER: Luchinat E 

PROVIDER: S-EPMC8489230 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature