title: Screening for small molecule modulators of Hsp70 chaperone activity using protein aggregation suppression assays: inhibition of the plasmodial chaperone PfHsp70-1.
abstract: Plasmodium falciparum heat shock protein 70 (PfHsp70-1) is thought to play an essential role in parasite survival and virulence in the human host, making it a potential antimalarial drug target. A malate dehydrogenase based aggregation suppression assay was adapted for the screening of small molecule modulators of Hsp70. A number of small molecules of natural (marine prenylated alkaloids and terrestrial plant naphthoquinones) and related synthetic origin were screened for their effects on the protein aggregation suppression activity of purified recombinant PfHsp70-1. Five compounds (malonganenone A-C, lapachol and bromo-β-lapachona) were found to inhibit the chaperone activity of PfHsp70-1 in a concentration dependent manner, with lapachol preferentially inhibiting PfHsp70-1 compared to another control Hsp70. Using growth inhibition assays on P. falciparum infected erythrocytes, all of the compounds, except for malonganenone B, were found to inhibit parasite growth with IC(50) values in the low micromolar range. Overall, this study has identified two novel classes of small molecule inhibitors of PfHsp70-1, one representing a new class of antiplasmodial compounds (malonganenones). In addition to demonstrating the validity of PfHsp70-1 as a possible drug target, the compounds reported in this study will be potentially useful as molecular probes for fundamental studies on Hsp70 chaperone function.
authors.affiliation: Biomedical Biotechnology Research Unit, Department of Biochemistry, Microbiology and Biotechnology, Rhodes University, Grahamstown, South Africa.
authors.forename: Ingrid L
authors.initials: IL
authors.lastname: Cockburn
authors.forename: Eva-Rachele
authors.initials: ER
authors.lastname: Pesce
authors.forename: Jude M
authors.initials: JM
authors.lastname: Pryzborski
authors.forename: Michael T
authors.initials: MT
authors.lastname: Davies-Coleman
authors.forename: Peter G K
authors.initials: PG
authors.lastname: Clark
authors.forename: Robert A
authors.initials: RA
authors.lastname: Keyzers
authors.forename: Linda L
authors.initials: LL
authors.lastname: Stephens
authors.forename: Gregory L
authors.initials: GL
authors.lastname: Blatch
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D000962:Antimalarials / Q000494:pharmacology*; D050884:HSP72 Heat-Shock Proteins / Q000187:drug effects / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D009285:Naphthoquinones / Q000494:pharmacology; D010963:Plasmodium falciparum / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000470:Alkaloids; D000962:Antimalarials; D050884:HSP72 Heat-Shock Proteins; D009285:Naphthoquinones; C008252:lapachol