FLUORESCENTLY-LABELED F-ACTIN PROTEIN BIOSENSORS AND METHODS OF HIGH-THROUGHPUT DRUG DISCOVERY

摘要

Protein biosensors for drug discovery, such as a high-throughput assay utilizing TR-F to detect the properties of binding proteins (e.g., actin-binding proteins, ABPs) using a fluorescence lifetime plate reader and fluorescent probes. In particular, the present invention is a new use of TR-F technique to rapidly evaluate binding of F-actin (- or + Tm) with cMyBP-C in solution. Changes in labeled actin fluorescence lifetime due to cMyBP-C phosphorylation and/or HCM mutations correlated with binding measured by traditional cosedimentation. The present invention features methods for HTS for identifying molecules that modulate cMyBP-C or cMyBP-C-actin complex.