BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> 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;...

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Autores principales:	Cilly Bernardette Schnider, Damaris Bausch-Fluck, Francis Brühlmann, Volker T. Heussler, Paul-Christian Burda
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	BioID blood stage liver stage PVM Plasmodium membranes Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/93b9ceafeb7547fdae09a5111f21a513
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spelling	oai:doaj.org-article:93b9ceafeb7547fdae09a5111f21a5132021-11-15T15:22:02ZBioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane10.1128/mSphere.00522-172379-5042https://doaj.org/article/93b9ceafeb7547fdae09a5111f21a5132018-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00522-17https://doaj.org/toc/2379-5042ABSTRACT 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 systematic proteomic analysis of the protein content of the PVM has been difficult in the past, due to difficulties encountered in attempts 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 PVM-resident exported protein 1 to biotinylate proteins of the PVM. These we further processed by affinity purification, liquid chromatography-tandem 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 (PlasmoDB accession no. PBANKA_1328000) and two conserved Plasmodium proteins with unknown functions (PBANKA_0519300 and PBANKA_0509000). In conclusion, our report 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. IMPORTANCE Intracellular pathogens are often surrounded by a host-cell derived membrane. This membrane is modified by the pathogens to their own needs and is crucial for their intracellular lifestyle. In Plasmodium parasites, this membrane is referred to as the PVM and only a limited number of its proteins are known so far. Here, we applied in rodent P. berghei parasites a method called BioID, which is based on biotinylation of proximal and interacting proteins by the promiscuous biotin ligase BirA*, and demonstrated its usefulness in identification of novel PVM proteins.Cilly Bernardette SchniderDamaris Bausch-FluckFrancis BrühlmannVolker T. HeusslerPaul-Christian BurdaAmerican Society for MicrobiologyarticleBioIDblood stageliver stagePVMPlasmodiummembranesMicrobiologyQR1-502ENmSphere, Vol 3, Iss 1 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	BioID blood stage liver stage PVM Plasmodium membranes Microbiology QR1-502
spellingShingle	BioID blood stage liver stage PVM Plasmodium membranes Microbiology QR1-502 Cilly Bernardette Schnider Damaris Bausch-Fluck Francis Brühlmann Volker T. Heussler Paul-Christian Burda BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane
description	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 systematic proteomic analysis of the protein content of the PVM has been difficult in the past, due to difficulties encountered in attempts 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 PVM-resident exported protein 1 to biotinylate proteins of the PVM. These we further processed by affinity purification, liquid chromatography-tandem 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 (PlasmoDB accession no. PBANKA_1328000) and two conserved Plasmodium proteins with unknown functions (PBANKA_0519300 and PBANKA_0509000). In conclusion, our report 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. IMPORTANCE Intracellular pathogens are often surrounded by a host-cell derived membrane. This membrane is modified by the pathogens to their own needs and is crucial for their intracellular lifestyle. In Plasmodium parasites, this membrane is referred to as the PVM and only a limited number of its proteins are known so far. Here, we applied in rodent P. berghei parasites a method called BioID, which is based on biotinylation of proximal and interacting proteins by the promiscuous biotin ligase BirA*, and demonstrated its usefulness in identification of novel PVM proteins.
format	article
author	Cilly Bernardette Schnider Damaris Bausch-Fluck Francis Brühlmann Volker T. Heussler Paul-Christian Burda
author_facet	Cilly Bernardette Schnider Damaris Bausch-Fluck Francis Brühlmann Volker T. Heussler Paul-Christian Burda
author_sort	Cilly Bernardette Schnider
title	BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane
title_short	BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane
title_full	BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane
title_fullStr	BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane
title_full_unstemmed	BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane
title_sort	bioid reveals novel proteins of the <italic toggle="yes">plasmodium</italic> parasitophorous vacuole membrane
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/93b9ceafeb7547fdae09a5111f21a513
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BioID Reveals Novel Proteins of the <italic toggle="yes">Plasmodium</italic> Parasitophorous Vacuole Membrane

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