Опубликована: Дек. 19, 2024

Deep Mutational Scanning (DMS) is an emerging method to systematically test the functional consequences of thousands sequence changes a protein target in single experiment. Because its utility interpreting both human variant effects and structure-function relationships, it holds substantial promise improve drug discovery clinical development. However, applications this domain require improved experimental analytical methods. To address need, we report novel DMS methods precisely quantitatively interrogate disease-relevant mechanisms, protein-ligand interactions, assess predicted response treatment. Using these methods, performed melanocortin-4 receptor (MC4R), G protein-coupled (GPCR) implicated obesity active development efforts. We assessed >6,600 amino acid substitutions on MC4R’s function across 18 distinct conditions, resulting >20 million unique measurements. From this, identified variants that have MC4R-mediated Gα s - q -signaling pathways, which could be used design drugs selectively bias activity. also pathogenic are likely amenable corrector therapy. Finally, functionally characterized structural relationships distinguish binding peptide versus small molecule ligands, guide compound optimization. Collectively, results demonstrate powerful empower

Язык: Английский