The long-term goal of this research is to design new compounds to inhibit UDP-galactopyranose mutase (UGM), a key enzyme in the biosynthesis of galactofuranose (Galf). Galf is an important component of the cell wall and glycan complexes found on the cell surface of many eukaryotic pathogens. It has been shown that UGM is essential for growth in helminths and is an important virulence factor in Aspergillus fumigatus and Leishmania major. Because of its importance in pathogenesis and its absence in humans, UGM is a target for the development of novel therapeutics against multiple eukaryotic pathogens. Previous studies by the PIs have determined that eukaryotic UGMs contain a unique ADP binding site and novel mechanistic role for the flavin cofactor. The goals of this project are to identify inhibitors of eukaryotic UGMs, characterize the molecular mechanisms of inhibition using biochemical and structural methods, and test the effects of these compounds on microbial and human cells. A subset of compounds having the highest potency and best safety profiles will be used as the starting structures for iterative optimization using structure-guided medicinal chemistry approaches to increase binding affinity, specificity, and drug-likeness. The team of investigators has expertise in UGM biochemistry and high-throughput screening (Sobrado), X-ray crystallography of UGM (Tanner), medicinal/synthetic organic chemistry (Gates), and toxicology/pharmacology (Ehrich). A major innovation of this project is that we direct high-throughput screening to the heretofore unexplored ADP pocket of eukaryotic UGMs. The molecular environment within this site is unique to and conserved by eukaryotic UGMs and, thus, the discovered compounds have the potential to inhibit multiple eukaryotic UGMs.
Infections by eukaryotic pathogens, such as fungi, parasites, and nematodes cause a multitude of diseases, including aspergillosis, Chagas? disease, leishmaniasis, elephantiasis, and river blindness. Despite the significant worldwide mortality and human suffering caused by these (mostly neglected) diseases, there are no efficacious drug treatments or vaccinations available to treat them. This project will discover inhibitors of UDP- galactopyranose mutase, the central enzyme in the biosynthesis of galactofuranose, as a first step toward the development of novel therapeutics for eukaryotic pathogens.
| Pierdominici-Sottile, Gustavo; Cossio-Pérez, Rodrigo; Da Fonseca, Isabel et al. (2018) Steric Control of the Rate-Limiting Step of UDP-Galactopyranose Mutase. Biochemistry 57:3713-3721 |
| Martín Del Campo, Julia S; Eckshtain-Levi, Meital; Sobrado, Pablo (2017) Identification of eukaryotic UDP-galactopyranose mutase inhibitors using the ThermoFAD assay. Biochem Biophys Res Commun 493:58-63 |
| Martin Del Campo, Julia S; Eckshtain-Levi, Meital; Vogelaar, Nancy J et al. (2017) Identification of Aspergillus fumigatus UDP-Galactopyranose Mutase Inhibitors. Sci Rep 7:10836 |
| Sobrado, Pablo; Tanner, John J (2017) Multiple functionalities of reduced flavin in the non-redox reaction catalyzed by UDP-galactopyranose mutase. Arch Biochem Biophys 632:59-65 |
| Martín Del Campo, Julia S; Vogelaar, Nancy; Tolani, Karishma et al. (2016) Inhibition of the Flavin-Dependent Monooxygenase Siderophore A (SidA) Blocks Siderophore Biosynthesis and Aspergillus fumigatus Growth. ACS Chem Biol 11:3035-3042 |
| Mehra-Chaudhary, Ritcha; Dai, Yumin; Sobrado, Pablo et al. (2016) In Crystallo Capture of a Covalent Intermediate in the UDP-Galactopyranose Mutase Reaction. Biochemistry 55:833-6 |
| Da Fonseca, Isabel; Qureshi, Insaf A; Mehra-Chaudhary, Ritcha et al. (2014) Contributions of unique active site residues of eukaryotic UDP-galactopyranose mutases to substrate recognition and active site dynamics. Biochemistry 53:7794-804 |
| Tanner, John J; Boechi, Leonardo; Andrew McCammon, J et al. (2014) Structure, mechanism, and dynamics of UDP-galactopyranose mutase. Arch Biochem Biophys 544:128-41 |
| Fonseca, Isabel O; Kizjakina, Karina; Sobrado, Pablo (2013) UDP-galactopyranose mutases from Leishmania species that cause visceral and cutaneous leishmaniasis. Arch Biochem Biophys 538:103-10 |
| Boechi, Leonardo; de Oliveira, Cesar Augusto F; Da Fonseca, Isabel et al. (2013) Substrate-dependent dynamics of UDP-galactopyranose mutase: Implications for drug design. Protein Sci 22:1490-501 |
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