Development of an online p38α mitogen-activated protein kinase binding assay and integration of LC-HR-MS.

ABSTRACT: A high-resolution screening method was developed for the p38α mitogen-activated protein kinase to detect and identify small-molecule binders. Its central role in inflammatory diseases makes this enzyme a very important drug target. The setup integrates separation by high-performance liquid chromatography with two parallel detection techniques. High-resolution mass spectrometry gives structural information to identify small molecules while an online enzyme binding detection method provides data on p38α binding. The separation step allows the individual assessment of compounds in a mixture and links affinity and structure information via the retention time. Enzyme binding detection was achieved with a competitive binding assay based on fluorescence enhancement which has a simple principle, i

SUBMITTER: Falck D 

PROVIDER: S-EPMC2943585 | biostudies-literature | 2010 Oct

REPOSITORIES: biostudies-literature