Use of Affimer Technology to Detect Conformational Changes in Protein Structure

Abstract: As the number one cause of death worldwide, cardiovascular disease (CVD) due to chronic inflammation is a major health concern. The increased inflammatory activity associated with atherosclerosis, a major cause of CVD, is partially due to increased levels of pro-inflammatory cytokines, particularly interleukin-6 (IL-6). Suppressor of cytokine signalling (SOCS) 3 protein is an important down regulator of the IL-6 signalling pathway and consequently of atherosclerotic plaque formation.
The actions of the universal second messenger cAMP are mediated by two sensors: PKA and, the more recently identified, exchange protein directly activated by cAMP (EPAC). In vascular endothelial cells (VECs), EPAC1 is a key mediator of SOCS3 induction as well as a central controller of anti-inflammatory processes. The overall aim of my project is therefore to identify EPAC1 modulators with the purpose of upregulating SOCS3 protein expression in VECs with the view of preventing inflammation associated with atherosclerosis. This will be done specifically using Affimers: an engineered non-antibody scaffold. Using this Affimer technology will allow us to develop novel Affimer-based compound screens to characterise agonist- and antagonist- conformational changes of EPAC1.