Antibiotic resistance is a significant threat to human health in all parts of the world. New resistance mechanisms are emerging and spreading globally, threatening our ability to treat common infectious diseases and necessitating new strategies for developing antimicrobial compounds. Moonlighting proteins are a key contributor to bacterial virulence. These proteins are multifunctional proteins in which a single protein performs multiple independent functions in different cell compartments, often making use of different conformations to do so. Moonlighting proteins that act as virulence factors tend to play key roles in conserved metabolic processes and elicit relatively muted responses from the immune system. Their evolutionary conservation, evasion of immune response and critical role in pathogenesis suggest that moonlighting proteins may make excellent targets for therapeutic development. One way to target these moonlighting proteins is with nanobodies. Nanobodies are single-domain fragments generated from camelid antibodies that combine the beneficial properties of small molecules and monoclonal antibodies, thus making them an appealing agent for creating new antibiotics. Nanobodies are easier to produce cheaply in bulk than antibodies while maintaining excellent thermal and pH stability. Just as importantly, nanobodies may be developed using a combination of surface display and directed evolution methods allowing for expedited drug discovery in response to the development of resistance. This poster outlines progress toward the expression and purification of moonlighting proteins from S. aureus and S. epidermidis for use in the directed evolution of next-generation antibiotics.
Albion College Foundation for Undergraduate Research, Scholarship, and Creative Activity Albion College Faculty Development Fund
