Project 464451

Yeast species of the Candida genus are the primary cause of opportunistic infections that represent a major public health problem. The spectrum of diseases caused by Candida species ranges from vaginal infections, to life-threatening bloodstream infections. Our long-term goal is to provide basic scientific knowledge about Candida cellular processes that are essential for pathogenesis and infectivity, to be used in development of novel anti-fungal therapies. In this current application, we propose to develop small molecule inhibitors for enzymatic activities within the shikimate pathway. This seven-step pathway is essential for Candida's viability and is absent in animals and humans, which positions its enzymes as particularly attractive targets for novel antimicrobial therapies. The shikimate pathway in Candida is mediated by a large protein called Aro1 that contains five distinct domains. Our structural and functional analysis has demonstrated that only four of the five C. albicans domains are functional. Even more importantly, our analysis has demonstrated that inactivation of each of these four functional Aro1 domains incapacitates the growth of C. albicans, providing an opportunity for inhibition through small molecules as segue for development of novel targeted antifungal therapeutics. As a proof of concept, we completed a screen which identified hit compounds capable of inhibiting the shikimate kinase domain of C. albicans Aro1. We also developed activity assays for each of the four essential Aro1 domains, setting the stage for further inhibitor screening. We also obtained high resolution molecular 3D images of Aro1 enzymatic domains, which will guide optimization of hit compounds towards higher potency and specificity. We will leverage this new molecular data to discover and advance a repertoire of small molecules that can target C. albicans Aro1 essential domains to serve as leads for further anti-fungal drug development.