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Proteomic analysis and studies of interactions of exported proteins in Plasmodium falciparum

English title Proteomic analysis and studies of interactions of exported proteins in Plasmodium falciparum
Applicant Beck Hans-Peter
Number 132709
Funding scheme Project funding (Div. I-III)
Research institution Swiss Tropical and Public Health Institute
Institution of higher education University of Basel - BS
Main discipline Molecular Biology
Start/End 01.10.2010 - 30.09.2013
Approved amount 415'000.00
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Keywords (12)

MALARIA; PLASMODIUM FALCIPARUM; PFEMP1; trafficking; exported proteins; protein interaction; MAHRP1; MAHRP2; Tethers; virulence factor; protein trafficking,; interactions

Lay Summary (English)

Lead
Lay summary
Plasmodium falciparum, the causative agent of malaria tropica infects human red blood cells. This process is unique because the erythrocyte is a highly specialized cell lacking organelles to support growing and dividing parasites. All metabolic pathways and developmental requirements must be installed by the parasite during the 48 h cycle within a red blood cell. In addition, the parasite substantially modifies the host cell resulting in an adhesive cell and thus leading to substantial morbidity. These cell modifications include the transport of the major virulence factor to the surface of the infected cell. However, invading such a highly specialized cell is rare in biology and requires the interplay of a specific and unique set of parasite proteins to refurbish these cells. In this project, we aim to elucidate the interaction and function of proteins which are exported early during the life cycle from the parasite into the host cell. For this we will use recombinant transfection technology, proteomic analysis, and other molecular techniques to identify and understand essential processes in the parasite's quest to survive in the human host cell. Only the understanding of these processes might enable us to design innovative intervention strategies against this deadly disease.
Direct link to Lay Summary Last update: 21.02.2013

Responsible applicant and co-applicants

Employees

Collaboration

Group / person Country
Types of collaboration
University of Oxford Great Britain and Northern Ireland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
Bio21 Australia (Oceania)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Research Infrastructure
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
BioMalPar meeting 14.05.2012 Heidelberg, Germany
Molecular Approaches to Malaria 19.02.2012 Lorne, Australia
BioMalPar meeting 16.05.2011 Heidelberg, Germany


Associated projects

Number Title Start Funding scheme
149297 Establishment of an interaction network of exported proteins in Plasmodium falciparum 01.10.2013 Project funding (Div. I-III)
149896 Mode of action of antimalarial synthetic peroxides 01.10.2013 Project funding (Div. I-III)
118456 Sequence and expression analysis of Plasmodium falciparum var genes and immunological and functional analysis of PfEMP1 in naturally infected human blood samples 01.10.2007 Project funding (Div. I-III)

Abstract

Background and Hypothesis:Plasmodium falciparum, the causative agent of malaria tropica invades erythrocytes and this stage is the only stage responsible for pathology. Invasion into an empty cell such as the erythrocyte requires unique processes immediately after invasion. These processes involve a complete restructuring of the cell, establishment of a protein translation and trafficking machinery to subsequently deposit adhesion conferring ligands onto the surface of the erythrocyte. We are interested in the interplay of parasite derived proteins which occur early in the erythrocyte life cycle and which are translocated into the cytosol of the infected erythrocyte. We and others have previously shown that some of these proteins are essential in the trafficking of the virulence determinants and that some of these proteins are not amendable to gene knock out approaches, suggesting their necessity for the survival of the parasite. In this project we would like to elucidate the protein-protein interactions of these early exported proteins. Objectives and approach:The overall objective is to elucidate the function and the identification of interaction partners of a group of previously identified PEXEL-negative exported proteins in P. falciparum.In detail we want toI) further develop our existing transfection platform to generate tagged versions of identified proteins to elucidate their localization and to identify interaction partners using pull down experiments and immuno-precipitations.II) generate knock out parasite lines for phenotypical analysis, or if impossible to establish conditional knock down parasite lines using the FKBP-DD approach.III) establish a recombinant screening system for interaction partners using a membrane bound split ubiquitin system in yeast or using a classical yeast two hybrid system for soluble proteins.IV) establish the proteome of tethers by differential ultracentrifugation and mass-spectrometry.V) analyse changes in the proteomic pattern upon over-expression or deletion of genes both for protein expression levels but also for post-translational protein modifications applying whole proteome fluorescent 2D-DIGE (differential gel electrophoresis) with subsequent mass-spectrometrical analysis. Expected outcome:We this project we hope to understand early events essential for parasite survival upon invasion into its host cell. This will not only deepen our understanding of the biology of P. falciparum but might provide new targets for innovative interventions.
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