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HeLa Based Cell Free Expression Systems for Expression of Plasmodium Rhoptry Proteins
Published on: June 10, 2015
Plasmodium falciparum Rab1A Localizes to Rhoptries in Schizonts
David Morse1, Wesley Webster1, Ming Kalanon1
1School of BioSciences, University of Melbourne, Melbourne, VIC, 3010, Australia.
Abstract:
Over-expression of a GFP-PfRab1A fusion protein in Plasmodium falciparum schizonts produces a punctate pattern of fluorescence typical of rhoptries, secretory organelles involved in host cell invasion. The GFP-positive bodies were purified by a combination of differential and density gradient centrifugation and their protein content determined by MS/MS sequencing. Consistent with the GFP rhoptry-like pattern of transgenic parasites, four of the 19 proteins identified have been previously described to be rhoptry-associated and another four are ER or ER-associated proteins. Confirmation that GFP-PfRab1A decorates rhoptries was obtained by its co-localization with Rap1 and Ron4 in late phase schizonts. We conclude that PfRab1A potentially regulates vesicular traffic from the endoplasmic reticulum to the rhoptries in Apicomplexa parasites.
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