Probing unfolding barriers in high throughput

Abstract

A protein's primary sequence encodes its energy landscape, including the thermodynamic stability and dynamics of all accessible conformations. While the field has come a long way in understanding how thermodynamics are encoded, how the sequence encodes kinetic barriers is still poorly understood. These kinetic barriers are important as they determine the rate at which a folded protein makes excursion to and from any unfolded and potentially harmful states. Therefore, we have developed a novel combination of yeast display and proteolysis to select for high kinetic barriers to unfolding.