BioID reveals novel proteins of the Plasmodium parasitophorous vacuole membrane

ABSTRACT: During their development within the vertebrate host, Plasmodium parasites infect hepatocytes and red blood cells. Within these cells, parasites are surrounded by a parasitophorous vacuole membrane (PVM). The PVM plays an essential role for the interaction of parasites with their host cells, however only a limited number of proteins of this membrane have been identified so far. This is partially because a systematic proteomic analysis of the PVM’s protein content has been difficult in the past, due to difficulties to separate the PVM from other membranes such as the parasite plasma membrane. In this study, we adapted the BioID technique to in vitro cultivated Plasmodium berghei blood stage parasites and utilized the promiscuous biotin ligase BirA* fused to the PVM-resident exported protein 1 to biotinylate proteins of the PVM. These we further processed by affinity purification, liquid chromatography based mass spectrometry (LC-MS/MS) and label-free quantitation leading to a list of 61 known and candidate PVM proteins. Seven proteins were analyzed further during blood and liver stage development. This resulted in the identification of three novel PVM proteins, which were the serine/threonine protein phosphatase UIS2 and two conserved Plasmodium proteins with unknown functions (PBANKA_0519300 and PBANKA_0509000). In conclusion, our study expands the number of known PVM proteins and experimentally validates BioID as a powerful method to screen for novel constituents of specific cellular compartments in P. berghei.

INSTRUMENT(S):

ORGANISM(S): Plasmodium Berghei Anka

TISSUE(S): Blood Cell, Blood Plasma

DISEASE(S): Plasmodium Falciparum Malaria

SUBMITTER: Damaris Bausch-Fluck  

LAB HEAD: Prof. Dr. Volker Heussler

PROVIDER: PXD007857 | Pride | 2018-01-03

REPOSITORIES: Pride