Discovery/Signaling Core

Thomas Jefferson University, Philadelphia PA

Investigators

Abstract

Core A - Abstract The Discovery/Screening Core provides high throughput screening (HTS), cryo-electron microscopy, signaling analysis, and virtual screening infrastructure to supply projects 1-4 with small molecules and mechanistic insight to exploit novel molecular mechanisms to promote bronchodilation in constrictive airway diseases. Conventional and computational screening components synergize to focus screening efforts in optimal regions of chemical space, generate and analyze structure-activity relationships (SAR), design and optimize screening workflows and disseminate the results and materials back to the scientific projects for detailed study and translational applications. The integration of conventional and virtual screening workflows will enable us to move rapidly from primary screening hits to detailed SAR, and from computational predictions to quantitative experimental data. We will further characterize biased allosteric modulators for the 2-adrenergic receptor (2AR) that we have previously identified and use HTS to identify new allosteric modulators (project 1). HTS will also be used to identify positive allosteric modulators (PAMs) for GPRC5A as well as negative allosteric modulators (NAMs) and antagonists for GPRC5B (project 4). We will use molecular modeling, flexible receptor docking and molecular dynamics simulations to explore the mechanisms of allosteric modulator binding to the 2AR (project 1), evaluate predicted all-atom phospholipase C complexes with protein kinase C (project 3), and model allosteric modulator binding to GPRC5A and GPRC5B and identify new candidates by virtual screening (project 4). All four projects will also be supported with specific reagents and signaling analysis including measurement of cAMP production, Ca2+ mobilization, -arrestin recruitment, GPCR-G protein coupling and GPCR internalization. Overall, the Discovery/Screening Core will generate data, lead compounds, biochemical probes, signaling analysis and mechanistic insight that will advance the aims of the scientific projects while developing innovative experimental and computational methodology that will enhance probe, drug and target discovery.