Dataset Information

Rapid proximity biotinylation of the Plasmodium falciparum exported protein, SBP1

ABSTRACT: The deadly human malaria-causing parasite, Plasmodium falciparum relies on its capacity to completely remodel its host red blood cell (RBC) through the export of several hundred parasite proteins across several membranes to the RBC.We fused the exported membrane protein, skeleton binding protein 1 (SBP1), with the rapid, efficient, promiscuous biotin ligase known as TurboID (SBP1TbID). Using time-resolved, proximity biotinylation and label-free quantitative proteomics, we identified early (pre-export) interactors and late (post-export) interactors of SBP1TbID.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Plasmodium Falciparum Nf54 (ncbitaxon:5843)

SUBMITTER: Vasant Muralidharan

PROVIDER: MSV000089743 | MassIVE | Mon Jun 27 17:05:00 BST 2022

SECONDARY ACCESSION(S): PXD034946

REPOSITORIES: MassIVE

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Publications

Time-resolved proximity biotinylation implicates a porin protein in export of transmembrane malaria parasite effectors.

Anaguano David D Dedkhad Watcharatip W Brooks Carrie F CF Cobb David W DW Muralidharan Vasant V

Journal of cell science 20231018 20

The malaria-causing parasite, Plasmodium falciparum completely remodels its host red blood cell (RBC) through the export of several hundred parasite proteins, including transmembrane proteins, across multiple membranes to the RBC. However, the process by which these exported membrane proteins are extracted from the parasite plasma membrane for export remains unknown. To address this question, we fused the exported membrane protein, skeleton binding protein 1 (SBP1), with TurboID, a rapid, effici ...[more]

PMID: 37772444

Similar Datasets

Project description:Toxoplasma gondii is a widely prevalent parasite of animals and humans that causes a broad spectrum of conditions ranging from asymptomatic or mild cold-like infections to abortion, severe ocular disease, encephalitis, and sometimes death. It is a member of the phylum Apicomplexa that also includes such significant human pathogens as Plasmodium spp. and Cryptosporidium spp. causing malaria and diarrheal disease, respectively. The toxoplasma cell has a multilayered pellicle: under the plasma membrane lies the inner membrane complex (IMC) comprised of flat membrane sacks supported by cortical microtubules and a meshwork of cytoskeletal filaments. The IMC acts as a mechanical barrier for membrane vesicles trafficking to and from the plasma membrane, necessitating the existence of dedicated cites for endo- and exocytosis where the IMC is interrupted. One such site is located at the apical end of the cell where the contents of the apical secretory organelles is exocytosed upon invasion. However, endocytosis in Toxoplasma and other apicomplexans is poorly understood, although conspicuous invaginations of the plasma membrane observed microscopically have been dubbed the micropore and postulated to be involved in endocytosis. We found that the endocytic markers including the subunits α and μ of the AP2 adaptor complex, dynamin-related protein C (DrpC), an Eps15 homology domain-containing protein, and the kelch-domain protein K13 localise to distinct persistent foci in the pellicle of Toxoplasma. Another type of persistent punctate structures found in the IMC are the enigmatic apical annuli whose function is completely unknown. We have discovered a new component of the annuli which is a putative LMBR1 family region protein embedded into the plasma membrane. Our experiments indicate that this protein – and annuli by extension – has a role in exocytosis. To better understand the molecular composition of these endocytic and exocytic structures and gain an insight into their function, we genetically fused them with a promiscuous bacterial biotin-ligase BirA* and used them as baits in proximity-dependent biotin identification experiments (BioID). We employed TMT10plex labelling and MS3-level tandem mass spectrometry with synchronous precursor selection (SPS-MS3) to achieve accurate quantification of multiple samples. Several hundred proteins have been identified and quantified in three biological replicates for every bait and the parental cell line control. Proteins significantly enriched in the BirA*-tagged cells relative to the control have been identified using linear modelling with LIMMA.