Dynamically-resolved structural studies of proteins from fungal and bacterial pathogens

Hauptman-Woodward Medical Research Inst, Buffalo NY

Investigators

Abstract

Summary Drug resistance and desensitization is a growing global concern. To overcome these therapeutic issues, drug discovery pipelines have to make use of all available structural information to generate novel, potent and safe therapeutics. Current medicinal chemistry efforts typically make use of static, cryo-temperature protein structures, which do not integrate the dynamics known govern protein function. Advancing from structure-activity to dynamic-activity relationships will diversify the way we pursue drug discovery, opening doors to more advanced therapeutics. To obtain this knowledge, the projects proposed will pursue time-resolved structural studies of medically-relevant proteins from fungal and bacterial pathogens. Although challenging, time- resolved structural biology has matured over the last decade making it amenable to a wider range of protein targets. This research will break methodological barriers to swiftly obtain dynamic- activity relationships, from which we will determine how dynamics correlate to protein function. Complementary biochemical and biophysical studies will be used to unambiguously resolve protein metastable intermediates. Methods such as photocaging and cryo-trapping are used to trigger synchronize protein function in the sample, making use of the work and expertise of the PI as well as the available facilities, including a dedicated organic synthesis laboratory. Two initial targets will be pursued. The first one, C. Auris 3-Deoxy-D-arabinoheptulosonate 7- phosphate Synthase (DAHPS) is an enzyme essential for fungal survival as it catalysis a step in the biosynthesis of aromatic amino-acids. It is considered a promising drug target to current antifungals and it is dynamically allosterically regulated. The second target is rubrerythrin from B. pseudomallei, a protein believed to be responsible for oxidative stress responses for which the mechanism is still unclear. Both projects will be targeted using an overarching methodological pipeline which will generate new insights into dynamic-activity relationships and provide structures of short-lived intermediates. Beyond the cutting-edge work proposed in this grant, these discoveries will fuel a long-term research program focusing on harnessing dynamic information to drive novel drug design. The fundamental work proposed in this grant will fill gaps in our knowledge of protein mechanism and its relationship to dynamics, and will pave the way to novel drug discovery approaches. The proposed time-resolved experiments are challenging but made possible by the multi-disciplinary expertise of the PI as well as strong collaborations that drive fertile scientific discussions and unobstructed access to cutting edge technology and facilities in the USA and abroad.