Journal article
VAR2CSA Ectodomain Labeling in Plasmodium falciparum Infected Red Blood Cells and Analysis via Flow Cytometry
OMS Carmo, MWA Dixon
Bio Protocol | BIO-PROTOCOL | Published : 2023
Abstract
Presentation of the variant antigen Plasmodium falciparum erythrocyte membrane protein 1 (EMP1) at the surface of infected red blood cells (RBCs) underpins the malaria parasite's pathogenicity. The transport of EMP1 to the RBC surface is facilitated by a parasite-derived trafficking system, in which over 500 parasite proteins are exported into the host cell cytoplasm. To understand how genetic ablation of selected exported proteins affects EMP1 transport, several EMP1 surface presentation assays have been developed, including: 1) trypsinization of surface-exposed EMP1 and analysis by SDS-PAGE and immunoblotting; and 2) infected RBC binding assays, to determine binding efficiency to immobiliz..
View full abstractGrants
Awarded by National Health and Medical Research Council
Funding Acknowledgements
We thank Michael Duffy and Stephen Rogerson for providing us with critical reagents. This original protocol was based on Beeson et al. (2004) and Smith et al. (1995) . We also thank Molly P. Schneider for reviewing the protocol and for her helpful suggestions. MWAD thanks the National Health and Medical Research Council (1098992) (https:// www.nhmrc.gov.au) for funding this work. The funders had no role in study design, data collection and analysis, the decision to publish, or the preparation of the manuscript.