Home Selective modulation of plasmodial Hsp70s by small molecules with antimalarial activity
Article
Licensed
Unlicensed Requires Authentication

Selective modulation of plasmodial Hsp70s by small molecules with antimalarial activity

  • Ingrid L. Cockburn , Aileen Boshoff , Eva-Rachele Pesce and Gregory L. Blatch EMAIL logo
Published/Copyright: April 18, 2014
Biological Chemistry
From the journal Biological Chemistry Volume 395 Issue 11

Abstract

Plasmodial heat shock protein 70 (Hsp70) chaperones represent a promising new class of antimalarial drug targets because of the important roles they play in the survival and pathogenesis of the malaria parasite Plasmodium falciparum. This study assessed a set of small molecules (lapachol, bromo-β-lapachona and malonganenones A, B and C) as potential modulators of two biologically important plasmodial Hsp70s, the parasite-resident PfHsp70-1 and the exported PfHsp70-x. Compounds of interest were assessed for modulatory effects on the steady-state basal and heat shock protein 40 (Hsp40)-stimulated ATPase activities of PfHsp70-1, PfHsp70-x and human Hsp70, as well as on the protein aggregation suppression activity of PfHsp70-x. The antimalarial marine alkaloid malonganenone A was of particular interest, as it was found to have limited cytotoxicity to mammalian cell lines and exhibited the desired properties of an effective plasmodial Hsp70 modulator. This compound was found to inhibit plasmodial and not human Hsp70 ATPase activity (Hsp40-stimulated), and hindered the aggregation suppression activity of PfHsp70-x. Furthermore, malonganenone A was shown to disrupt the interaction between PfHsp70-x and Hsp40. This is the first report to show that PfHsp70-x has chaperone activity, is stimulated by Hsp40 and can be specifically modulated by small molecule compounds.

Keywords: heat shock proteins; malonganenones; naphthoquinones; PfHsp70-1; PfHsp70-x; Plasmodium falciparum
Corresponding author: Gregory L. Blatch, Biomedical Biotechnology Research Unit (BioBRU), Department of Biochemistry and Microbiology, Rhodes University, Grahamstown 6140, South Africa; and College of Health and Biomedicine, Victoria University, Melbourne 8001, Victoria, Australia, e-mail:

Acknowledgments

Many thanks to Prof. M.T. Davies Coleman and Dr. R. Keyzers for supplying the compounds included in this study, and thanks to Dr. J. de la Mare and Dr. E. Prinsloo for assistance with mammalian cell culture and SPR experiments, respectively. This research was funded by a Deutsche Forschungsgemeinschaft (DFG) German-African Cooperation Project in Infectology grant [DFG (Ref: LI 402/12-0)] as well as by the National Research Foundation (NRF, South Africa). ILC was funded by Rhodes University and the NRF.

References

Acharya, P., Kumar, R., and Tatu, U. (2007). Chaperoning a cellular upheaval in malaria: heat shock proteins in Plasmodium falciparum. Mol. Biochem. Parasitol. 153, 85–94.10.1016/j.molbiopara.2007.01.009Search in Google Scholar PubMed

Anderson, T. (2009). Mapping the spread of malaria drug resistance. PLoS Med 6, e1000054.10.1371/journal.pmed.1000054Search in Google Scholar PubMed PubMed Central

Assimon, V.A., Gillies, A.T., Rauch, J.N., and Gestwicki, J.E. (2013). Hsp70 protein complexes as drug targets. Curr. Pharm. Des. 19, 404–417.10.2174/138161213804143699Search in Google Scholar PubMed PubMed Central

Balaburski, G.M., Leu, J.I.-J., Beeharry, N., Hayik, S., Andrake, M.D., Zhang, G., Herlyn, M., Villanueva, J., Dunbrack, R.L., Yen, T., et al. (2013). A modified HSP70 inhibitor shows broad activity as an anticancer agent. Mol. Cancer Res. 11, 219–229.10.1158/1541-7786.MCR-12-0547-TSearch in Google Scholar PubMed PubMed Central

Banerjee, T., Singh, R.R., Gupta, S., Surolia, A., and Surolia, N. (2012). 15-Deoxyspergualin hinders physical interaction between basic residues of transit peptide in PfENR and Hsp70-1. IUBMB Life 64, 99–107.10.1002/iub.583Search in Google Scholar PubMed

Bell, S.L., Chiang, A.N., and Brodsky, J.L. (2011). Expression of a malarial Hsp70 improves defects in chaperone-dependent activities in ssa1 mutant yeast. PloS One 6, e20047.10.1371/journal.pone.0020047Search in Google Scholar PubMed PubMed Central

Botha, M., Chiang, A.N., Needham, P.G., Stephens, L.L., Hoppe, H.C., Külzer, S., Przyborski, J.M., Lingelbach, K., Wipf, P., Brodsky, J.L., et al. (2011). Plasmodium falciparum encodes a single cytosolic type I Hsp40 that functionally interacts with Hsp70 and is upregulated by heat shock. Cell Stress Chaperon 16, 389–401.10.1007/s12192-010-0250-6Search in Google Scholar PubMed PubMed Central

Brodsky, J.L. and Chiosis, G. (2006). Hsp70 molecular chaperones: emerging roles in human disease and identification of small molecule modulators. Curr. Top. Med. Chem. 6, 1215–1225.10.2174/156802606777811997Search in Google Scholar PubMed

Cesa, L.C., Patury, S., Komiyama, T., Ahmad, A., Zuiderweg, E.R.P., and Gestwicki, J.E. (2013). Inhibitors of difficult protein-protein interactions identified by high-throughput screening of multiprotein complexes. ACS Chem. Biol. 8, 1988–1997.10.1021/cb400356mSearch in Google Scholar PubMed PubMed Central

Cheetham, M.E., Jackson, A.P., and Anderton, B.H. (1994). Regulation of 70-kDa heat-shock-protein ATPase activity and substrate binding by human DnaJ-like proteins, HSJ1a and HSJ1b. Eur. J. Biochem. 226, 99–107.10.1111/j.1432-1033.1994.tb20030.xSearch in Google Scholar PubMed

Chiang, A.N., Valderramos, J.-C., Balachandran, R., Chovatiya, R.J., Mead, B.P., Schneider, C., Bell, S.L., Klein, M.G., Huryn, D.M., Chen, X.S., et al. (2009). Select pyrimidinones inhibit the propagation of the malarial parasite, Plasmodium falciparum. Bioorg. Med. Chem. 17, 1527–1533.10.1016/j.bmc.2009.01.024Search in Google Scholar

Clark, I.A., Chaudhri, G., and Cowden, W.B. (1989). Some roles of free radicals in malaria. Free Radic. Biol. Med. 6, 315–321.10.1016/0891-5849(89)90058-0Search in Google Scholar

Cockburn, I.L., Pesce, E.-R., Pryzborski, J.M., Davies-Coleman, M.T., Clark, P.G.K., Keyzers, R.A., Stephens, L.L., and Blatch, G.L. (2011). Screening for small molecule modulators of Hsp70 chaperone activity using protein aggregation suppression assays: inhibition of the plasmodial chaperone PfHsp70-1. Biol. Chem. 392, 431–438.10.1515/bc.2011.040Search in Google Scholar

Craig, A. and Scherf, A. (2001). Molecules on the surface of the Plasmodium falciparum infected erythrocyte and their role in malaria pathogenesis and immune evasion. Mol. Biochem. Parasitol. 115, 129–143.10.1016/S0166-6851(01)00275-4Search in Google Scholar

De Koning-Ward, T.F., Gilson, P.R., Boddey, J.A., Rug, M., Smith, B.J., Papenfuss, A.T., Sanders, P.R., Lundie, R.J., Maier, A.G., Cowman, A.F., et al. (2009). A newly discovered protein export machine in malaria parasites. Nature 459, 945–949.10.1038/nature08104Search in Google Scholar PubMed PubMed Central

Dondorp, A.M., Nosten, F., Yi, P., Das, D., Phyo, A.P., Tarning, J., Lwin, K.M., Ariey, F., Hanpithakpong, W., Lee, S.J., et al. (2009). Artemisinin resistance in Plasmodium falciparum malaria. N. Engl. J. Med. 361, 455–467.10.1056/NEJMoa0808859Search in Google Scholar PubMed PubMed Central

Evans, C.G., Chang, L., and Gestwicki, J.E. (2010). Heat shock protein 70 (Hsp70) as an emerging drug target. J. Med. Chem. 53, 4585–4602.10.1021/jm100054fSearch in Google Scholar PubMed PubMed Central

Fewell, S.W., Smith, C.M., Lyon, M.A., Dumitrescu, T.P., Wipf, P., Day, B.W., and Brodsky, J.L. (2004). Small molecule modulators of endogenous and co-chaperone-stimulated Hsp70 ATPase activity. J. Biol. Chem. 279, 51131–51140.10.1074/jbc.M404857200Search in Google Scholar PubMed

Gao, X.-C., Zhou, C.-J., Zhou, Z.-R., Wu, M., Cao, C.-Y., and Hu, H.-Y. (2012). The C-terminal helices of heat shock protein 70 are essential for J-domain binding and ATPase activation. J. Biol. Chem. 287, 6044–6052.10.1074/jbc.M111.294728Search in Google Scholar PubMed PubMed Central

Grover, M., Chaubey, S., Ranade, S., and Tatu, U. (2013). Identification of an exported heat shock protein 70 in Plasmodium falciparum. Parasite 20, 2.10.1051/parasite/2012002Search in Google Scholar PubMed PubMed Central

Haldar, K. and Mohandas, N. (2007). Erythrocyte remodeling by malaria parasites. Curr. Opin. Hematol. 14, 203–209.10.1097/MOH.0b013e3280f31b2dSearch in Google Scholar

Hatherley, R., Blatch, G.L., and Bishop, O.T. (2013). Plasmodium falciparum Hsp70-x: a heat shock protein at the host-parasite interface. J. Biomol. Struct. Dyn. doi: 10.1080/07391102.2013.834849 [Epub ahead of print].10.1080/07391102.2013.834849Search in Google Scholar

Hennessy, F., Nicoll, W.S., Zimmermann, R., Cheetham, M.E., and Blatch, G.L. (2005). Not all J domains are created equal: Implications for the specificity of Hsp40-Hsp70 interactions. Protein Sci. Publ. Protein Soc. 14, 1697–1709.10.1110/ps.051406805Search in Google Scholar

Jinwal, U.K., Koren, J., O’Leary, J.C., Jones, J.R., Abisambra, J.F., and Dickey, C.A. (2010). Hsp70 ATPase modulators as therapeutics for Alzheimer’s and other neurodegenerative diseases. Mol. Cell. Pharmacol. 2, 43–46.Search in Google Scholar

Johnson, D.S., Weerapana, E., and Cravatt, B.F. (2010). Strategies for discovering and derisking covalent, irreversible enzyme inhibitors. Future Med. Chem. 2, 949–964.10.4155/fmc.10.21Search in Google Scholar

Joshi, B., Biswas, S., and Sharma, Y.D. (1992). Effect of heat-shock on Plasmodium falciparum viability, growth and expression of the heat-shock protein “PFHSP70-I” gene. FEBS Lett. 312, 91–94.10.1016/0014-5793(92)81417-KSearch in Google Scholar

Kampinga, H.H. and Craig, E.A. (2010). The HSP70 chaperone machinery: J proteins as drivers of functional specificity. Nat. Rev. Mol. Cell Biol. 11, 750–750.10.1038/nrm2982Search in Google Scholar

Külzer, S., Rug, M., Brinkmann, K., Cannon, P., Cowman, A., Lingelbach, K., Blatch, G.L., Maier, A.G., and Przyborski, J.M. (2010). Parasite-encoded Hsp40 proteins define novel mobile structures in the cytosol of the P. falciparum-infected erythrocyte. Cell. Microbiol. 12, 1398–1420.10.1111/j.1462-5822.2010.01477.xSearch in Google Scholar

Külzer, S., Charnaud, S., Dagan, T., Riedel, J., Mandal, P., Pesce, E.R., Blatch, G.L., Crabb, B.S., Gilson, P.R., and Przyborski, J.M. (2012). Plasmodium falciparum-encoded exported hsp70/hsp40 chaperone/co-chaperone complexes within the host erythrocyte. Cell. Microbiol. 14, 1784–1795.10.1111/j.1462-5822.2012.01840.xSearch in Google Scholar

Kumar, N., Koski, G., Harada, M., Aikawa, M., and Zheng, H. (1991). Induction and localization of Plasmodium falciparum stress proteins related to the heat shock protein 70 family. Mol. Biochem. Parasitol. 48, 47–58.10.1016/0166-6851(91)90163-ZSearch in Google Scholar

Kumar, R., Musiyenko, A., and Barik, S. (2003). The heat shock protein 90 of Plasmodium falciparum and antimalarial activity of its inhibitor, geldanamycin. Malar. J. 2, 30.10.1186/1475-2875-2-30Search in Google Scholar PubMed PubMed Central

Li, X., Srinivasan, S.R., Connarn, J., Ahmad, A., Young, Z.T., Kabza, A.M., Zuiderweg, E.R.P., Sun, D., and Gestwicki, J.E. (2013). Analogues of the allosteric heat shock protein 70 (Hsp70) inhibitor, MKT-077, as anti-cancer agents. ACS Med. Chem. Lett. 130917101701007.10.1021/ml400204nSearch in Google Scholar PubMed PubMed Central

Massey, A.J. (2010). ATPases as drug targets: insights from heat shock proteins 70 and 90. J. Med. Chem. 53, 7280–7286.10.1021/jm100342zSearch in Google Scholar PubMed

Matambo, T.S., Odunuga, O.O., Boshoff, A., and Blatch, G.L. (2004). Overproduction, purification, and characterization of the Plasmodium falciparum heat shock protein 70. Protein Expr. Purif. 33, 214–222.10.1016/j.pep.2003.09.010Search in Google Scholar PubMed

Mayer, M.P. and Bukau, B. (2005). Hsp70 chaperones: cellular functions and molecular mechanism. Cell. Mol. Life Sci. 62, 670–684.10.1007/s00018-004-4464-6Search in Google Scholar PubMed PubMed Central

Misra, G. and Ramachandran, R. (2009). Hsp70-1 from Plasmodium falciparum: protein stability, domain analysis and chaperone activity. Biophys. Chem. 142, 55–64.10.1016/j.bpc.2009.03.006Search in Google Scholar PubMed

Miyata, Y., Rauch, J.N., Jinwal, U.K., Thompson, A.D., Srinivasan, S., Dickey, C.A., and Gestwicki, J.E. (2012). Cysteine reactivity distinguishes redox sensing by the heat-inducible and constitutive forms of heat shock protein 70. Chem. Biol. 19, 1391–1399.10.1016/j.chembiol.2012.07.026Search in Google Scholar PubMed PubMed Central

Nyalwidhe, J. and Lingelbach, K. (2006). Proteases and chaperones are the most abundant proteins in the parasitophorous vacuole of Plasmodium falciparum-infected erythrocytes. Proteomics 6, 1563–1573.10.1002/pmic.200500379Search in Google Scholar PubMed

Oakley, M.S.M., Kumar, S., Anantharaman, V., Zheng, H., Mahajan, B., Haynes, J.D., Moch, J.K., Fairhurst, R., McCutchan, T.F., and Aravind, L. (2007). Molecular factors and biochemical pathways induced by febrile temperature in intraerythrocytic Plasmodium falciparum parasites. Infect. Immun. 75, 2012–2025.10.1128/IAI.01236-06Search in Google Scholar PubMed PubMed Central

Pavithra, S.R., Banumathy, G., Joy, O., Singh, V., and Tatu, U. (2004). Recurrent fever promotes Plasmodium falciparum development in human erythrocytes. J. Biol. Chem. 279, 46692–46699.10.1074/jbc.M409165200Search in Google Scholar PubMed

Pesce, E.-R., Acharya, P., Tatu, U., Nicoll, W.S., Shonhai, A., Hoppe, H.C., and Blatch, G.L. (2008). The Plasmodium falciparum heat shock protein 40, Pfj4, associates with heat shock protein 70 and shows similar heat induction and localisation patterns. Int. J. Biochem. Cell Biol. 40, 2914–2926.10.1016/j.biocel.2008.06.011Search in Google Scholar PubMed

Ramya, T.N.C., Surolia, N., and Surolia, A. (2006). 15-Deoxyspergualin modulates Plasmodium falciparum heat shock protein function. Biochem. Biophys. Res. Commun. 348, 585–592.10.1016/j.bbrc.2006.07.082Search in Google Scholar

Sargeant, T.J., Marti, M., Caler, E., Carlton, J.M., Simpson, K., Speed, T.P., and Cowman, A.F. (2006). Lineage-specific expansion of proteins exported to erythrocytes in malaria parasites. Genome Biol. 7, R12.10.1186/gb-2006-7-2-r12Search in Google Scholar

Sharma, Y.D. (1992). Structure and possible function of heat-shock proteins in Falciparum malaria. Comp. Biochem. Physiol. B 102, 437–444.10.1016/0305-0491(92)90033-NSearch in Google Scholar

Shonhai, A., Boshoff, A., and Blatch, G.L. (2005). Plasmodium falciparum heat shock protein 70 is able to suppress the thermosensitivity of an Escherichia coli DnaK mutant strain. Mol. Genet. Genomics MGG 274, 70–78.10.1007/s00438-005-1150-9Search in Google Scholar PubMed

Shonhai, A., Boshoff, A., and Blatch, G.L. (2007). The structural and functional diversity of Hsp70 proteins from Plasmodium falciparum. Protein Sci. 16, 1803–1818.10.1110/ps.072918107Search in Google Scholar PubMed PubMed Central

Shonhai, A., Botha, M., de Beer, T.A.P., Boshoff, A., and Blatch, G.L. (2008). Structure-function study of a Plasmodium falciparum Hsp70 using three dimensional modelling and in vitro analyses. Protein Pept. Lett. 15, 1117–1125.10.2174/092986608786071067Search in Google Scholar PubMed

Szabo, A., Langer, T., Schroder, H., Flanagan, J., Bukau, B., and Hartl, F.U. (1994). The ATP hydrolysis-dependent reaction cycle of the Escherichia coli Hsp70 system DnaK, DnaJ, and GrpE. Proc. Natl. Acad. Sci. USA 91, 10345–10349.10.1073/pnas.91.22.10345Search in Google Scholar PubMed PubMed Central

Wellems, T.E. and Plowe, C.V. (2001). Chloroquine-Resistant malaria. J. Infect. Dis. 184, 770–776.10.1086/322858Search in Google Scholar PubMed

Witkowski, B., Lelièvre, J., Barragán, M.J.L., Laurent, V., Su, X., Berry, A., and Benoit-Vical, F. (2010). Increased tolerance to artemisinin in Plasmodium falciparum is mediated by a quiescence mechanism. Antimicrob. Agents Chemother. 54, 1872–1877.10.1128/AAC.01636-09Search in Google Scholar PubMed PubMed Central

Wright, C.M., Chovatiya, R.J., Jameson, N.E., Turner, D.M., Zhu, G., Werner, S., Huryn, D.M., Pipas, J.M., Day, B.W., Wipf, P., et al. (2008). Pyrimidinone-peptoid hybrid molecules with distinct effects on molecular chaperone function and cell proliferation. Bioorg. Med. Chem. 16, 3291–3301.10.1016/j.bmc.2007.12.014Search in Google Scholar PubMed PubMed Central

World Health Organization (2012). World Malaria Report 2012. Geneva: World Health Organization.10.30875/ac42f7b8-enSearch in Google Scholar

Supplemental Material

The online version of this article (DOI: 10.1515/hsz-2014-0138) offers supplementary material, available to authorized users.

Received: 2014-2-15
Accepted: 2014-4-12
Published Online: 2014-4-18
Published in Print: 2014-11-1

©2014 by De Gruyter

Articles in the same Issue

  1. Frontmatter
  2. Guest Editorial
  3. Integrating Epigenetics
  4. HIGHLIGHT: NEW INSIGHTS IN EPIGENETICS
  5. Epigenetic control of hematopoiesis: the PU.1 chromatin connection
  6. Role of lncRNAs in prostate cancer development and progression
  7. Polycomb and Trithorax group protein-mediated control of stress responses in plants
  8. Transcription as a force partitioning the eukaryotic genome
  9. The epigenetic tracks of aging
  10. Early epigenetic cancer decisions
  11. Reviews
  12. Functions of the neuron-specific protein ADAP1 (centaurin-α1) in neuronal differentiation and neurodegenerative diseases, with an overview of structural and biochemical properties of ADAP1
  13. Titin: central player of hypertrophic signaling and sarcomeric protein quality control
  14. Research Articles/Short Communications
  15. Protein Structure and Function
  16. Selective modulation of plasmodial Hsp70s by small molecules with antimalarial activity
https://doi.org/10.1515/hsz-2014-0138
Keywords for this article
heat shock proteins; malonganenones; naphthoquinones; PfHsp70-1; PfHsp70-x; Plasmodium falciparum

Articles in the same Issue

  1. Frontmatter
  2. Guest Editorial
  3. Integrating Epigenetics
  4. HIGHLIGHT: NEW INSIGHTS IN EPIGENETICS
  5. Epigenetic control of hematopoiesis: the PU.1 chromatin connection
  6. Role of lncRNAs in prostate cancer development and progression
  7. Polycomb and Trithorax group protein-mediated control of stress responses in plants
  8. Transcription as a force partitioning the eukaryotic genome
  9. The epigenetic tracks of aging
  10. Early epigenetic cancer decisions
  11. Reviews
  12. Functions of the neuron-specific protein ADAP1 (centaurin-α1) in neuronal differentiation and neurodegenerative diseases, with an overview of structural and biochemical properties of ADAP1
  13. Titin: central player of hypertrophic signaling and sarcomeric protein quality control
  14. Research Articles/Short Communications
  15. Protein Structure and Function
  16. Selective modulation of plasmodial Hsp70s by small molecules with antimalarial activity
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 9.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/hsz-2014-0138/html
Scroll to top button