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Background  Recent work has perfected yeast-based methods for measuring drug transport by the  Plasmodium falciparum  chloroquine (CQ) resistance transporter (PfCRT).    Methods  The approach relies on inducible heterologous expression of PfCRT in  Saccharomyces cerevisiae  yeast. In these experiments selecting drug concentrations are not toxic to the yeast, nor is expression of PfCRT alone toxic. Only when PfCRT is expressed in the presence of CQ is the growth of yeast impaired, due to inward transport of chloroquine (CQ) via the transporter.    Results  During analysis of all 53 known naturally occurring PfCRT isoforms, two isoforms (PH1 and PH2 PfCRT) were found to be intrinsically toxic to yeast, even in the absence of CQ. Additional analysis of six very recently identified PfCRT isoforms from Malaysia also showed some toxicity. In this paper the nature of this yeast toxicity is examined. Data also show that PH1 and PH2 isoforms of PfCRT transport CQ with an efficiency intermediate to that catalyzed by previously studied CQR conferring isoforms. Mutation of PfCRT at position 160 is found to perturb vacuolar physiology, suggesting a fitness cost to position 160 amino acid substitutions.    Conclusion  These data further define the wide range of activities that exist for PfCRT isoforms found in  P. falciparum  isolates from around the globe.   Graphical abstract   V5 Western blot quantifying relative PfCRT expression in total (CM) vs vacuolar (VAC) yeast membranes for unmodified HB3 PfCRT vs HB3 and PH1 PMA–PfCRT chimeras.

Plasmodium falciparum chloroquine resistance transporter (PfCRT) isoforms PH1 and PH2 perturb vacuolar physiology