Chemically Mapping Malarial Rupture Proteases

Summary

Principal Investigator: Matthew Bogyo
Abstract: Malaria is a devastating disease that causes significant mortality in many countries of the developing world. The most deadly form of the disease is caused by the opportunistic pathogen, Plasmodium falciparum. Significant efforts have been made to understand the process by which the parasite invades a host cell to establish infection, yet relatively little is known about the process by which the parasite mediates its release after replication has occurred. This process is essential for propagation of the pathogen and agents that block rupture are likely to be valuable for development as novel anti-malarial agents. This proposal outlines plans to use small molecules to study the functional roles of proteases that regulate the process of host cell rupture. Specifically, it describes the use of phenotypic screens using libraries of protease inhibitors to identify compounds that block the release of parasites from host red blood cells. Screening hits will be used to identify protease targets and to dissect the details of their regulation of host cell rupture. Finally, lead compounds will be applied to mouse models of malaria to validate multiple proteases as drug targets for novel anti-malarial therapies. PUBLIC HEALTH RELEVANCE: This project outlines plans to use small molecules to identify proteases used by the parasite pathogen, Plasmodium falciparum, to mediate rupture of host red blood cells during the blood stage infection of a human host. This work will lead to the identification of potentially valuable targets for development of new therapeutic treatment strategies for malaria.
Funding Period: 2009-02-01 - 2015-01-31
more information: NIH RePORT

Top Publications

  1. pmc Small molecule-induced allosteric activation of the Vibrio cholerae RTX cysteine protease domain
    Patrick J Lupardus
    Department of Molecular and Cellular Physiology and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Science 322:265-8. 2008
  2. pmc Functional imaging of proteases: recent advances in the design and application of substrate-based and activity-based probes
    Laura E Edgington
    Cancer Biology Program, Stanford University School of Medicine, 300 Pasteur Dr, Stanford, CA 94305 5324, USA
    Curr Opin Chem Biol 15:798-805. 2011
  3. pmc Proteomic analysis of fractionated Toxoplasma oocysts reveals clues to their environmental resistance
    Heather M Fritz
    Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California Davis, Davis, California, United States of America
    PLoS ONE 7:e29955. 2012
  4. pmc A nonpeptidic cathepsin S activity-based probe for noninvasive optical imaging of tumor-associated macrophages
    Martijn Verdoes
    Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Chem Biol 19:619-28. 2012
  5. pmc Caspase-1 activity is required to bypass macrophage apoptosis upon Salmonella infection
    Aaron W Puri
    Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, USA
    Nat Chem Biol 8:745-7. 2012
  6. pmc Validation of the proteasome as a therapeutic target in Plasmodium using an epoxyketone inhibitor with parasite-specific toxicity
    Hao Li
    Graduate Program in Chemical and Systems Biology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305 5324, USA
    Chem Biol 19:1535-45. 2012
  7. pmc The antimalarial natural product symplostatin 4 is a nanomolar inhibitor of the food vacuole falcipains
    Sara Christina Stolze
    Zentrum für Medizinische Biotechnologie, Fakultat fur Biologie, Universitat Duisburg Essen, Universitätsstr 2, 45117 Essen, Germany
    Chem Biol 19:1546-55. 2012
  8. pmc Bradyzoite pseudokinase 1 is crucial for efficient oral infectivity of the Toxoplasma gondii tissue cyst
    Kerry R Buchholz
    Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA
    Eukaryot Cell 12:399-410. 2013
  9. pmc Target deconvolution techniques in modern phenotypic profiling
    Jiyoun Lee
    Department of Global Medical Science, Sungshin Women s University, Seoul 142 732, Republic of Korea
    Curr Opin Chem Biol 17:118-26. 2013
  10. pmc Small-molecule inhibition of a depalmitoylase enhances Toxoplasma host-cell invasion
    Matthew A Child
    Department of Pathology, Stanford University School of Medicine, Stanford, California, USA
    Nat Chem Biol 9:651-6. 2013

Research Grants

Detail Information

Publications28

  1. pmc Small molecule-induced allosteric activation of the Vibrio cholerae RTX cysteine protease domain
    Patrick J Lupardus
    Department of Molecular and Cellular Physiology and Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Science 322:265-8. 2008
    ..Biochemical and kinetic analyses of CPD mutants indicate that InsP6 binding induces an allosteric switch that leads to the autoprocessing and intracellular release of toxin-effector domains...
  2. pmc Functional imaging of proteases: recent advances in the design and application of substrate-based and activity-based probes
    Laura E Edgington
    Cancer Biology Program, Stanford University School of Medicine, 300 Pasteur Dr, Stanford, CA 94305 5324, USA
    Curr Opin Chem Biol 15:798-805. 2011
    ..In particular, it highlights the strengths and weaknesses of both substrate-based and activity-based probes and their applications for imaging cysteine proteases that are important biomarkers for multiple human diseases...
  3. pmc Proteomic analysis of fractionated Toxoplasma oocysts reveals clues to their environmental resistance
    Heather M Fritz
    Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California Davis, Davis, California, United States of America
    PLoS ONE 7:e29955. 2012
    ..The latter are known from other systems to be key to enabling survival against desiccation...
  4. pmc A nonpeptidic cathepsin S activity-based probe for noninvasive optical imaging of tumor-associated macrophages
    Martijn Verdoes
    Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Chem Biol 19:619-28. 2012
    ..Thus, BMV083 is a potential valuable in vivo reporter for tumor-associated macrophages that could greatly facilitate the future studies of macrophage function in the process of tumorigenesis...
  5. pmc Caspase-1 activity is required to bypass macrophage apoptosis upon Salmonella infection
    Aaron W Puri
    Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, USA
    Nat Chem Biol 8:745-7. 2012
    ..Furthermore, we report that upon Salmonella infection, inflammasome-mediated caspase-1 activity is required to bypass apoptosis in favor of proinflammatory pyroptotic cell death...
  6. pmc Validation of the proteasome as a therapeutic target in Plasmodium using an epoxyketone inhibitor with parasite-specific toxicity
    Hao Li
    Graduate Program in Chemical and Systems Biology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305 5324, USA
    Chem Biol 19:1535-45. 2012
    ..Subsequently, we used PR3 to significantly reduce parasite load in Plasmodium berghei infected mice without host toxicity, thus validating the proteasome as a viable antimalarial drug target...
  7. pmc The antimalarial natural product symplostatin 4 is a nanomolar inhibitor of the food vacuole falcipains
    Sara Christina Stolze
    Zentrum für Medizinische Biotechnologie, Fakultat fur Biologie, Universitat Duisburg Essen, Universitätsstr 2, 45117 Essen, Germany
    Chem Biol 19:1546-55. 2012
    ....
  8. pmc Bradyzoite pseudokinase 1 is crucial for efficient oral infectivity of the Toxoplasma gondii tissue cyst
    Kerry R Buchholz
    Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, USA
    Eukaryot Cell 12:399-410. 2013
    ..Together, these data reveal that BPK1 plays a crucial role in the in vivo development and infectivity of Toxoplasma cysts...
  9. pmc Target deconvolution techniques in modern phenotypic profiling
    Jiyoun Lee
    Department of Global Medical Science, Sungshin Women s University, Seoul 142 732, Republic of Korea
    Curr Opin Chem Biol 17:118-26. 2013
    ..In this review, we will outline how both new and old techniques are being used in the difficult process of target identification and validation as well as discuss some of the ongoing challenges remaining for phenotypic screening...
  10. pmc Small-molecule inhibition of a depalmitoylase enhances Toxoplasma host-cell invasion
    Matthew A Child
    Department of Pathology, Stanford University School of Medicine, Stanford, California, USA
    Nat Chem Biol 9:651-6. 2013
    ..We demonstrate that TgPPT1 is a bona fide depalmitoylase, thereby establishing an important role for dynamic and reversible palmitoylation in host-cell invasion by T. gondii. ..
  11. pmc Applications of small molecule probes in dissecting mechanisms of bacterial virulence and host responses
    Aaron W Puri
    Department of Chemical and Systems Biology, Department of Microbiology and Immunology, and Department of Pathology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, California 94305, United States
    Biochemistry 52:5985-96. 2013
    ..This review describes the development and application of activity-based probes for examining aspects of bacterial infection on both sides of the host-pathogen interface. ..
  12. pmc Improved quenched fluorescent probe for imaging of cysteine cathepsin activity
    Martijn Verdoes
    Departments of Pathology, Cancer Biology Program, and Microbiology and Immunology, Stanford School of Medicine, 300 Pasteur Drive, Stanford, California 94305 5324, United States
    J Am Chem Soc 135:14726-30. 2013
    ..These reagents show enhanced in vivo properties and broad reactivity resulting in dramatically improved labeling and tumor imaging properties compared to those of previously reported ABPs. ..
  13. pmc Ferrous iron-dependent drug delivery enables controlled and selective release of therapeutic agents in vivo
    Edgar Deu
    Departments of Pathology and Microbiology and Immunology, Stanford School of Medicine, Stanford, CA 94305
    Proc Natl Acad Sci U S A 110:18244-9. 2013
    ..This approach may find useful application in parasitic infections and more broadly in any disease state characterized by aberrant production of reactive ferrous iron. ..
  14. pmc Development of small molecule inhibitors and probes of human SUMO deconjugating proteases
    Victoria E Albrow
    Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Chem Biol 18:722-32. 2011
    ..The AOMK compound therefore represents promising new reagents to study the process of SUMO deconjugation...
  15. pmc Functional characterization of a SUMO deconjugating protease of Plasmodium falciparum using newly identified small molecule inhibitors
    Elizabeth L Ponder
    Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Chem Biol 18:711-21. 2011
    ..The most potent lead compound inhibited both recombinant PfSENP1 activity and P. falciparum replication in infected human blood. These studies provide valuable new tools for the study of SUMOylation in P. falciparum...
  16. pmc Using small molecules to dissect mechanisms of microbial pathogenesis
    Aaron W Puri
    Department of Chemical and Systems Biology, Stanford UniversitySchool of Medicine, 300 Pasteur Dr, Stanford, California 94305, USA
    ACS Chem Biol 4:603-16. 2009
    ..In this Review we highlight ways in which these techniques have been applied to examine bacterial and parasitic pathogenesis and discuss possible ways in which these efforts can be expanded in the near future...
  17. pmc Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag
    Aimee Shen
    Department of Pathology, Stanford School of Medicine, Stanford, California, United States of America
    PLoS ONE 4:e8119. 2009
    ..Furthermore, in addition to being timesaving, versatile, and inexpensive, our results indicate that the CPD purification system can enhance the expression, integrity, and solubility of intractable proteins from diverse organisms...
  18. pmc 4-Bromophenacyl bromide specifically inhibits rhoptry secretion during Toxoplasma invasion
    Sandeep Ravindran
    Department of Microbiology and Immunology, Stanford University, Stanford, California, United States of America
    PLoS ONE 4:e8143. 2009
    ..This potent compound, the modified "click-chemistry" forms of it, and the resistant mutants should serve as useful tools to further study the processes of Toxoplasma early invasion, in general, and rhoptry secretion, in particular...
  19. pmc Use of activity-based probes to develop high throughput screening assays that can be performed in complex cell extracts
    Edgar Deu
    Department of Pathology, Stanford School of Medicine, Stanford, California, United States of America
    PLoS ONE 5:e11985. 2010
    ..High throughput screening (HTS) is one of the primary tools used to identify novel enzyme inhibitors. However, its applicability is generally restricted to targets that can either be expressed recombinantly or purified in large quantities...
  20. pmc Functional studies of Plasmodium falciparum dipeptidyl aminopeptidase I using small molecule inhibitors and active site probes
    Edgar Deu
    Department of Pathology, Stanford School of Medicine, 300 Pasteur Drive, Stanford, CA 94305, USA
    Chem Biol 17:808-19. 2010
    ..Interestingly, we found that resynthesis and activation of DPAP1 after inhibition is rapid, suggesting that effective drugs would need to sustain DPAP1 inhibition for a period of 2-3 hr...
  21. pmc Biochemical characterization of Plasmodium falciparum dipeptidyl aminopeptidase 1
    Flora Wang
    Department of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA
    Mol Biochem Parasitol 175:10-20. 2011
    ..Two reversible cathepsin C inhibitors potently inhibited both recombinant and native DPAP1, thereby validating the use of recombinant DPAP1 for future inhibitor discovery and characterization...
  22. pmc Global profiling of proteolysis during rupture of Plasmodium falciparum from the host erythrocyte
    Paul W Bowyer
    Department of Pathology, Stanford University School of Medicine, Stanford, California 94305, USA
    Mol Cell Proteomics 10:M110.001636. 2011
    ..These data also provide insight into the biochemical events that take place during host cell rupture and are likely to be valuable for the study of proteases that could potentially be targeted for therapeutic gain...
  23. pmc Rational design of inhibitors and activity-based probes targeting Clostridium difficile virulence factor TcdB
    Aaron W Puri
    Department of Chemical and Systems Biology, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, California 94305, USA
    Chem Biol 17:1201-11. 2010
    ..The inhibitors presented will guide the development of therapeutics targeting C. difficile, and the probes will serve as tools for studying the unique activation mechanism of bacterial toxin CPDs...
  24. pmc Defining an allosteric circuit in the cysteine protease domain of Clostridium difficile toxins
    Aimee Shen
    Department of Pathology, Stanford School of Medicine, Stanford, CA, USA
    Nat Struct Mol Biol 18:364-71. 2011
    ..Collectively, our results identify an allosteric circuit that allows bacterial virulence factors to sense and respond to the eukaryotic environment...
  25. pmc Chemical genetic screen identifies Toxoplasma DJ-1 as a regulator of parasite secretion, attachment, and invasion
    Carolyn I Hall
    Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Proc Natl Acad Sci U S A 108:10568-73. 2011
    ..Together, our results suggest that TgDJ-1 plays an important role that is likely downstream of the calcium flux required for microneme secretion, parasite motility, and subsequent invasion of host cells...
  26. pmc Proteases as regulators of pathogenesis: examples from the Apicomplexa
    Hao Li
    Departments of Pathology and Microbiology and Immunology, Stanford University, Standford, CA 94305, USA
    Biochim Biophys Acta 1824:177-85. 2012
    ..This article is part of a Special Issue entitled: Proteolysis 50 years after the discovery of lysosome...
  27. pmc Chemical biology approaches for the study of apicomplexan parasites
    Matthew A Child
    Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA Electronic address
    Mol Biochem Parasitol 192:1-9. 2013
    ..As such, the parasite community has embraced them with great success. Some of these successes and the continuing evolution of chemical biology applied to apicomplexans will be discussed. ..

Research Grants30

  1. Mechanisms of mosquito midgut invasion by Plasmodium ookinetes
    Marcelo Jacobs-Lorena; Fiscal Year: 2013
    ..Knowledge generated by these studies may have important implications for the development of multivalent transmission-blocking vaccines. ..