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Infection and Immunity, July 2006, p. 4330-4338, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00054-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

A Set of Glycosylphosphatidyl Inositol-Anchored Membrane Proteins of Plasmodium falciparum Is Refractory to Genetic Deletion

Paul R. Sanders,1,2,3 Lev M. Kats,4 Damien R. Drew,1 Rebecca A. O'Donnell,1 Matthew O'Neill,1 Alexander G. Maier,1 Ross L. Coppel,4 and Brendan S. Crabb1*

Walter and Eliza Hall Institute of Medical Research, Melbourne, Australia,1 Cooperative Research Centre for Vaccine Technology, Brisbane, Australia,2 Department of Medical Biology, University of Melbourne, Melbourne, Australia,3 Department of Microbiology and the Victorian Bioinformatics Consortium, Monash University, Clayton, Victoria 3800, Australia4

Received 10 January 2006/ Returned for modification 5 March 2006/ Accepted 1 May 2006

Targeted gene disruption has proved to be a powerful approach for studying the function of important ligands involved in erythrocyte invasion by the extracellular merozoite form of the human malaria parasite, Plasmodium falciparum. Merozoite invasion proceeds via a number of seemingly independent alternate pathways, such that entry can proceed with parasites lacking particular ligand-receptor interactions. To date, most focus in this regard has been on single-pass (type 1) membrane proteins that reside in the secretory organelles. Another class of merozoite proteins likely to include ligands for erythrocyte receptors are the glycosylphosphatidyl inositol (GPI)-anchored membrane proteins that coat the parasite surface and/or reside in the apical organelles. Several of these are prominent vaccine candidates, although their functions remain unknown. Here, we systematically attempted to disrupt the genes encoding seven of the known GPI-anchored merozoite proteins of P. falciparum by using a double-crossover gene-targeting approach. Surprisingly, and in apparent contrast to other merozoite antigen classes, most of the genes (six of seven) encoding GPI-anchored merozoite proteins are refractory to genetic deletion, with the exception being the gene encoding merozoite surface protein 5 (MSP-5). No distinguishable growth rate or invasion pathway phenotype was detected for the msp-5 knockout line, although its presence as a surface-localized protein was confirmed.

* Corresponding author. Mailing address: Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC 3050, Australia. Phone: 61 3 9345 2555. Fax: 61 3 9347 0852. E-mail: crabb{at}wehi.edu.au.

Editor: J. F. Urban, Jr.

Infection and Immunity, July 2006, p. 4330-4338, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00054-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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