My laboratory uses approaches derived from both chemistry and biology. Projects include problems in inhibitor design, enzyme catalysis, protein degradation and the mechanism of visual disease. Techniques vary with the particular project, and can entail molecular biology, organic synthesis, protein crystallography and NMR spectroscopy as well as protein purification, enzyme kinetics and mutagenesis. Our ongoing projects are outlined below. For more information, please see the lab web site.
Dynamic structural determinants of reaction specificity. Understanding of how structure determines function is a central challenge in biochemistry. The IMPDH/GMPR family provides a striking example of how subtle differences in protein sequence, and hence in structure, can profoundly change reaction outcomes. These enzymes share a common set of catalytic residues and bind the same ligands with similar affinities. The reactions utilize the same covalent intermediate, yet with markedly different outcomes. IMPDH performs a hydride transfer reaction followed by a hydrolysis reaction, and a protein conformational changes rearranges the active site to accommodate both reactions. GMPR performs a deamination reaction followed by a hydride transfer reaction. In this case, the cofactor has a different position in each reaction. We are now trying to understand what structural features determine this very different dynamic behavior. Our long-range goal is to develop computational models that accurately recapitulate and quantitatively predict the catalytic properties of enzymes in the IMPDH/GMPR family. This work will advance the field of computational chemistry and provide important insights into how enzymes work. This work is a collaboration with Wei Yang of Florida State University.
Targeting a prokaryotic enzyme in a eukaryotic pathogen. The protozoan parasite Cryptosporidium parvum is an emerging opportunistic pathogen and potential bio-warfare agent. The C. parvum oocyte is resistant to the usual methods of water treatment, which has caused spectacular outbreaks such as the infection of 40% of the inhabitants of Milwaukee in 1993. C. parvum is resistant to the usual antiparasitic drugs and currently used chemotherapy is ineffective. In collaboration with Boris Striepen at UGA, we have been engaged in a medicinal chemistry program targeting C. parvum IMPDH. Curiously, the parasite obtained its IMPDH gene via horizontal transfer from a bacteria, so the parasite enzyme is very different from its host. We have a collection of low nanomolar inhibitors of the parasite enzyme, some of which show promising in vivo antiparasitic activity. These compounds also inhibit IMPDHs from pathogenic bacteria such as Mycobacterium tuberculosis, Staphylococcus aureus, Helicobacter pylori and Francisella tularensis. We are now investigating the potential of these compounds as broad spectrum antibiotics. This work is a collaboration with Joanna Goldberg at Emory and Barb Mann at UVA.
IMPeD: inhibitor mediated protein degradation. A formidable toolkit exists for manipulating protein expression at the transcriptional level, but the methods for post-translational modulation of proteins are few. A small molecule that induces degradation of endogenous proteins would clearly be a tremendously useful tool for probing protein function and an exciting new approach for chemotherapy. We serendipitously discovered a small molecule tag that induces the degradation of target proteins. We are currently investigating the mechanism of degradation and applying this method to clinically important targets.
Pathophysiological mechanisms of retinal disease. Many inherited retinal diseases are caused by mutations in proteins of the visual cycle, which can easily explain why disease is photoreceptor-specific. However, retinal disease can also result from mutations in widely expressed proteins. The photoreceptor-specific effects of these mutations are perplexing and pathophysiological mechanisms are undefined. One such protein is inosine monophosphate dehydrogenase type 1 (IMPDH1), which catalyzes a key step in guanine nucleotide biosynthesis. The effects of the IMPDH1 mutations cannot be explained by the loss of enzyme activity. We have recently discovered that IMPDH binds nucleic acids and demonstrated that the disease-causing mutations perturb nucleic acid binding. We are now investigating how this defect causes the specific apoptosis of photoreceptor cells with the aim of developing strategies for therapy.
Mechanistic enzymology. Filamentous fungi produce many important natural products such as penicillin and mycophenolic acid. New projects are available investigating the enzymes involved in these biosynthetic pathways.
Mortimer SE, Xu D, McGrew D, Hamaguchi N, Lim HC, Bowne SJ, Daiger SP, Hedstrom L. IMP dehydrogenase type 1 associates with polyribosomes translating rhodopsin mRNA. J Biol Chem. 2008 Dec 26;283(52):36354-60.
Hedstrom L, Liechti G, Goldberg JB, Gollapalli DR. The antibiotic potential of prokaryotic IMP dehydrogenase inhibitors. Curr Med Chem. 2011;18(13):1909-18.
Patton GC, Stenmark P, Gollapalli DR, Sevastik R, Kursula P, Flodin S, Schuler H, Swales CT, Eklund H, Himo F, Nordlund P, Hedstrom L. Cofactor mobility determines reaction outcome in the IMPDH and GMPR (beta-alpha)8 barrel enzymes. Nat Chem Biol. 2011;7(12):950-8.
Riera TV, Zheng L, Josephine HR, Min D, Yang W, Hedstrom L. Allosteric Activation via Kinetic Control: Potassium Accelerates a Conformational Change in IMP Dehydrogenase. Biochemistry. 2011;50(39):8508-18. PMCID: 3186055.
Sun XE, Hansen BG, Hedstrom L. Kinetically Controlled Drug Resistance: HOW PENICILLIUM BREVICOMPACTUM SURVIVES MYCOPHENOLIC ACID. J Biol Chem. 2011;286(47):40595-600. PMCID: 3220510.
Gorla SK, Kavitha M, Zhang M, Liu X, Sharling L, Gollapalli DR, Striepen B, Hedstrom L, Cuny GD. Selective and potent urea inhibitors of cryptosporidium parvum inosine 5'-monophosphate dehydrogenase. J Med Chem. 2012;55(17): 7759-71.
Hansen BG, Sun XE, Genee HJ, Kaas CS, Nielsen JB, Mortensen UH, Frisvad JC, Hedstrom L. Adaptive evolution of drug targets in producer and non-producer organisms. Biochem J. 2012;441(1):219-26.
Hedstrom L. The dynamic determinants of reaction specificity in the IMPDH/GMPR family of (beta/alpha)(8) barrel enzymes. Crit Rev Biochem Mol Biol. 2012;47(3):250-63. PMCID: 3337344.
Kirubakaran S, Gorla SK, Sharling L, Zhang M, Liu X, Ray SS, Macpherson IS, Striepen B, Hedstrom L, Cuny GD. Structure-activity relationship study of selective benzimidazole-based inhibitors of Cryptosporidium parvum IMPDH. Bioorg Med Chem Lett. 2012;22(5):1985-8. PMCID: 3289519.
Long MJ, Gollapalli DR, Hedstrom L. Inhibitor mediated protein degradation. Chem Biol. 2012;19(5):629-37. PMCID: 3361691.
McGrew DA, Hedstrom L. Towards a pathological mechanism for IMPDH1-linked retinitis pigmentosa. Adv Exp Med Biol.2012;723:539-45.
Johnson CR, Gorla SK, Kavitha M, Zhang M, Liu X, Striepen B, Mead JR, Cuny GD, Hedstrom L. Phthalazinone inhibitors of inosine-5'-monophosphate dehydrogenase from Cryptosporidium parvum. Bioorg Med Chem Lett. 2013; 23(4):1004-7. PMCID: 3557747.
Gorla SK, Kavitha M, Zhang M, Chin JE, Liu X, Striepen B, Makowska-Grzyska M, Kim Y, Joachimiak A, Hedstrom L, Cuny GD. Optimization of benzoxazole-based inhibitors of Cryptosporidium parvum inosine 5'-monophosphate dehydrogenase. J Med Chem. 2013 May 23;56(10):4028-43.
Mandapati K, Gorla SK, House AL, McKenney ES, Zhang M, Rao SN, Gollapalli DR, Mann BJ, Goldberg JB, Cuny GD, Glomski IJ, Hedstrom L. (2014). "Repurposing cryptosporidium inosine 5'-monophosphate dehydrogenase inhibitors as potential antibacterial agents." ACS Med Chem Lett 5(8): 846-850.
Gorla SK, McNair NN, Yang G, Gao S, Hu M, Jala VR, Haribabu B, Striepen B, Cuny GD, Mead JR, Hedstrom L. (2014). "Validation of IMP dehydrogenase inhibitors in a mouse model of cryptosporidiosis." Antimicrob Agents Chemother 58(3): 1603-1614.
Makowska-Grzyska M, Kim Y, Maltseva N, Osipiuk J, Gu M, Zhang M, Mandapati K, Gollapalli DR, Gorla SK, Hedstrom L, Joachimiak A. (2015). "A Novel Cofactor-binding Mode in Bacterial IMP Dehydrogenases Explains Inhibitor Selectivity." J Biol Chem 290(9): 5893-5911.
Makowska-Grzyska M, Kim Y, Gorla SK, Wei Y, Mandapati K, Zhang M, Maltseva N, Modi G, Boshoff HI, Gu M, Aldrich C, Cuny GD, Hedstrom L, Joachimiak A. (2015). "Mycobacterium tuberculosis IMPDH in Complexes with Substrates, Products and Antitubercular Compounds." PLoS ONE 10(10): e0138976.
Lawson AP, Long MJ, Coffey RT, Qian Y, Weerapana E, El Oualid F, Hedstrom L.. (2015). "Naturally Occurring Isothiocyanates Exert Anticancer Effects by Inhibiting Deubiquitinating Enzymes." Cancer Res 75(23): 5130-5142.
Hedstrom, L. (2015). "Cryptosporidium: a first step toward tractability." Trends Parasitol 31(9): 401-402.
Coffey RT, Shi Y, Long MJ, Marr MT 2nd, Hedstrom L. (2016). "Ubiquilin-mediated Small Molecule Inhibition of Mammalian Target of Rapamycin Complex 1 (mTORC1) Signaling." J Biol Chem 291(10): 5221-5233.
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Last update: August 2, 2016