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Antimalarial screening via large‐scale purification of P lasmodium falciparum C a 2+ ‐ ATP ase 6 and in vitro studies
Author(s) -
DavidBosne Stéphanie,
Florent Isabelle,
Lund Winther AnneMarie,
Hansen John B.,
BuchPedersen Morten,
Machillot Paul,
Maire Marc,
Jaxel Christine
Publication year - 2013
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.12244
Subject(s) - plasmodium falciparum , endoplasmic reticulum , atpase , in vitro , biology , serca , biochemistry , enzyme , pharmacology , chemistry , malaria , immunology
The most severe form of human malaria is caused by the parasite P lasmodium falciparum . Despite the current need, there is no effective vaccine and parasites are becoming resistant to most of the antimalarials available. Therefore, there is an urgent need to discover new drugs from targets that have not yet suffered from drug pressure with the aim of overcoming the problem of new emerging resistance. Membrane transporters, such as P . falciparum C a 2+ ‐ ATP ase 6 ( P f ATP 6), the P . falciparum sarcoplasmic/endoplasmic reticulum C a 2+ ‐ ATP ase ( SERCA ), have been proposed as potentially good antimalarial targets. The present investigation focuses on: (a) the large‐scale purification of P f ATP 6 for maintenance of its enzymatic activity; (b) screening for P f ATP 6 inhibitors from a compound library; and (c) the selection of the best inhibitors for further tests on P . falciparum growth in vitro . We managed to heterologously express in yeast and purify an active form of P f ATP 6 as previously described, although in larger amounts. In addition to some classical SERCA inhibitors, a chemical library of 1680 molecules was screened. From these, we selected a pool of the 20 most potent inhibitors of P f ATP 6, presenting half maximal inhibitory concentration values in the range 1–9 μ m . From these, eight were chosen for evaluation of their effect on P . falciparum growth in vitro , and the best compound presented a half maximal inhibitory concentration of ~ 2 μ m . We verified the absence of an inhibitory effect of most of the compounds on mammalian SERCA 1a, representing a potential advantage in terms of human toxicity. The present study describes a multidisciplinary approach allowing the selection of promising P f ATP 6‐specific inhibitors with good antimalarial activity.