Development of a high throughput cytochrome P450 ligand-binding assay.

ABSTRACT: Human cytochrome P450 enzymes are membrane-embedded monooxygenases responsible for xenobiotic metabolism, steroidogenesis, fatty acid metabolism, and vitamin metabolism. Their active sites can accommodate diverse small molecules and understanding these interactions is key to decoding enzymatic functionality and designing drugs. The most common method for characterizing small molecule binding is quantifying absorbance changes that typically occur when ligands enter the active site near the heme iron. Traditionally, such titrations are monitored by a spectrophotometer, requiring significant manual time, protein, and increasing solvents. This assay was adapted for semi-automated high throughput screening, increasing throughput 50-fold while requiring less protein and keeping solvent concentra

SUBMITTER: Frydendall EK 

PROVIDER: S-EPMC11530589 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature