Plasmodium falciparum PfEMP1 DiQ-BioID data

PfEMP1 is the major virulence factor of Plasmodium falciparum parasites. PfEMP1 is expressed as an integral membrane protein on the surface of infected red blood cells. As a part of this project, the BioID system was utilized to biotinylate proximal proteins of a functional PfEMP1. In order to do this, selection linked integration (SLI) was used to modify a specific PfEMP1 (PfIT_060021400; IT4var01) by fusing it with the promiscuous biotin ligase BirA* and obtain a parasite population predominantly expressing this modified variant. Three separate IT4 cell lines were generated where BirA* was fused to different positions of PfEMP1. In the first cell line (Position 1), BirA* is C-terminal to the ATS domain of PfEMP1, in the second cell line (position 2), BirA* is between the ATS and the transmembrane domain and in the third cell line (position 3), BirA* is between the transmembrane and the extracellular binding domains. Biotinylated proteins extracted with either triton or SDS containing buffers (using sequential extraction of the same parasite pellet) were isolated in large scales and subsequently analyzed via mass spectrometry in three separate experiments for the three BioID cell line as well as for IT4 control parasites (parent). The experiments resulted in a set of enriched proteins for each of the 3 cell lines (position 1-3) over IT4 parasites.