Regions of RAMA interacting with PfSortilin are sufficient for trafficking to the rhoptries. mCherry-RAMAC and mCherry-RAMAE fusion proteins traffic to the rhoptries, as confirmed by colocalization with RAP1, while RAMAA, RAMAB, and RAMAD fusions are trafficked to the parasitophorous vacuole (PV), the default destination for proteins harboring a signal sequence. We expressed and purified five different regions encompassing the whole protein (RAMAA to RAMAE), excluding the signal peptide and the GPI anchor sequence. These regions were incubated with parasite lysates from the PfSortilin-3HA-tagged line RAMAC contains 11 imperfect repeats of the sequence EE(S/KN). perhaps the RAMAC repeats are important for binding to PfSortilin. Examination of the transfectants showed that RAMAA, RAMAB, and RAMAD were found in the parasitophorous vacuole (PV), the default destination for proteins harboring a signal sequence (i, ii, and iv). In the case of RAMAC and RAMAE, a punctate pattern suggestive of an apical location was obtained (iii and v). Almost complete overlap between either RAMAC and RAMAE labeling with RAP1 by IFA confirmed their rhoptry localization (vi and vii).
Hallée S, Boddey JA, Cowman AF, Richard D. Evidence that the Plasmodium falciparum Protein Sortilin Potentially Acts as an Escorter for the Trafficking of the Rhoptry-Associated Membrane Antigen to the Rhoptries. mSphere. 2018 3(1). pii: e00551-17.
Other associated proteins
PFID | Formal Annotation |
---|---|
PF3D7_1410400 | rhoptry-associated protein 1 |
PF3D7_1451800 | sortilin |