Discovery of fragments inducing conformational effects in dynamic proteins using a second-harmonic generation biosensor.

ABSTRACT: Biophysical screening of compound libraries for the identification of ligands that interact with a protein is efficient, but does typically not reveal if (or how) ligands may interfere with its functional properties. For this a biochemical/functional assay is required. But for proteins whose function is dependent on a conformational change, such assays are typically complex or have low throughput. Here we have explored a high-throughput second-harmonic generation (SHG) biosensor to detect fragments that induce conformational changes upon binding to a protein in real time and identify dynamic regions. Multiwell plate format SHG assays were developed for wild-type and six engineered single-cysteine mutants of acetyl choline binding protein (AChBP), a homologue to ligand gated ion channels (L

SUBMITTER: FitzGerald EA 

PROVIDER: S-EPMC8694943 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature