Identification of Ligand Binding by Protein Stabilization: Comparison of ATLAS with Biophysical and Enzymatic Methods

• DOI: 10.1089/adt.2007.100
• Publication Date: 2008-03-12T21:04:02Z
• Authors: Peggy A. Thompson; Shaohui Wang; Lindsay J. Howett; Mei-Mei Wang; Rupal Patel; April Averill; Richard E. Showalter; Bin Li; James R. Appleman
• Keywords: Target protein

ATLAS (Any Target Ligand Affinity Screen) (Anadys Pharmaceuticals, Inc., San Diego, CA) is a homogeneous, affinity-based high-throughput screening technology based on protein thermal denaturation and the ability of ligands to bind stabilize target from unfolding. To further understand assay sensitivity for identification that soluble targets, firefly luciferase was chosen characterize technology. Luciferase multidomain with complex unfolding pathway. Binding ATP results in stabilizing conformational rearrangement domains. Using allowed us evaluate generality method ligand binding any target. inhibitors identified functional screens were used assess capability rank order inhibitors. Comparison 50% effective concentration other biophysical biochemical methods offered insight into optimizing conditions maximize compound stabilization. The show importance characterizing aggregation behavior allow screen be optimally designed.