Abstract
To enhance the repertoire of molecular tools for studying malaria parasite biology, we adapted a ligand-regulatable FKBP protein destabilization domain (ddFKBP) for use in P. falciparum. We destabilized the reporter yellow fluorescent protein (YFP) and the P. falciparum protease falcipain-2 in a ligand-reversible manner by tagging with ddFKBP. The swollen food vacuole phenotype of falcipain-2 knockout parasites could be rescued in a Shld1 ligand–dependent fashion by falcipain-2–ddFKBP expression.
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Acknowledgements
We thank I. Gluzman (Washington University) for help with transfections, E. Istvan (Washington University) for falcipain-2 knockout parasites, T. Wandless (Stanford University) for FKBP plasmids pBMN L106P-YFP iHcRed-t and pBMN YFP-L106P iHcRed-t as well as for Shld1 compound, Jacobus Pharmaceuticals for WR99210, M. Drew (Washington University) for microscopy advice, G. McFadden (University of Melbourne) for plasmids pCHDR-3/4 and PfHSP86 5′-pENTR4/1, MR4 and J. Adams (University of South Florida) for anti-BiP, and M. Meissner (University of Heidelburg) for helpful comments. This work was supported by US National Institutes of Health grant AI47798.
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C.M.A. designed and executed experiments and wrote the manuscript. D.E.G. designed experiments and wrote the manuscript.
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Armstrong, C., Goldberg, D. An FKBP destabilization domain modulates protein levels in Plasmodium falciparum. Nat Methods 4, 1007–1009 (2007). https://doi.org/10.1038/nmeth1132
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DOI: https://doi.org/10.1038/nmeth1132
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