title: Proteomic analysis reveals novel proteins associated with the Plasmodium protein exporter PTEX and a loss of complex stability upon truncation of the core PTEX component, PTEX150.
abstract: The Plasmodium translocon for exported proteins (PTEX) has been established as the machinery responsible for the translocation of all classes of exported proteins beyond the parasitophorous vacuolar membrane of the intraerythrocytic malaria parasite. Protein export, particularly in the asexual blood stage, is crucial for parasite survival as exported proteins are involved in remodelling the host cell, an essential process for nutrient uptake, waste removal and immune evasion. Here, we have truncated the conserved C-terminus of one of the essential PTEX components, PTEX150, in Plasmodium falciparum in an attempt to create mutants of reduced functionality. Parasites tolerated C-terminal truncations of up to 125 amino acids with no reduction in growth, protein export or the establishment of new permeability pathways. Quantitative proteomic approaches however revealed a decrease in other PTEX subunits associating with PTEX150 in truncation mutants, suggesting a role for the C-terminus of PTEX150 in regulating PTEX stability. Our analyses also reveal three previously unreported PTEX-associated proteins, namely PV1, Pf113 and Hsp70-x (respective PlasmoDB numbers; PF3D7_1129100, PF3D7_1420700 and PF3D7_0831700) and demonstrate that core PTEX proteins exist in various distinct multimeric forms outside the major complex.
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia.|Monash University, Melbourne, VIC, 3800, Australia.
authors.forename: Brendan
authors.initials: B
authors.lastname: Elsworth
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Sanders
authors.affiliation: Walter & Eliza Hall Institute, Melbourne, VIC, 3052, Australia.
authors.forename: Thomas
authors.initials: T
authors.lastname: Nebl
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, VIC, Australia.|ARC Centre of Excellence for Coherent X-ray Science, The University of Melbourne, Melbourne, VIC, Australia.|Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC, Australia.
authors.forename: Steven
authors.initials: S
authors.lastname: Batinovic
authors.affiliation: Deakin University, Waurn Ponds, 3216, Australia.
authors.forename: Ming
authors.initials: M
authors.lastname: Kalanon
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia.
authors.forename: Catherine Q
authors.initials: CQ
authors.lastname: Nie
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia.|Monash University, Melbourne, VIC, 3800, Australia.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Charnaud
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia.
authors.forename: Hayley E
authors.initials: HE
authors.lastname: Bullen
authors.affiliation: Deakin University, Waurn Ponds, 3216, Australia.
authors.forename: Tania F
authors.initials: TF
authors.lastname: de Koning Ward
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Melbourne, Melbourne, VIC, Australia.|ARC Centre of Excellence for Coherent X-ray Science, The University of Melbourne, Melbourne, VIC, Australia.|Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC, Australia.
authors.forename: Leann
authors.initials: L
authors.lastname: Tilley
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia.|Monash University, Melbourne, VIC, 3800, Australia.|University of Melbourne, Melbourne, VIC, 3010, Australia.
authors.forename: Brendan S
authors.initials: BS
authors.lastname: Crabb
authors.affiliation: Burnet Institute, 85 Commercial Road, Melbourne, VIC, 3004, Australia. gilson@burnet.edu.au.|Monash University, Melbourne, VIC, 3800, Australia. gilson@burnet.edu.au.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Gilson
mesh_terms: D002478:Cells, Cultured; D004912:Erythrocytes / Q000469:parasitology*; D006801:Humans; D026901:Membrane Transport Proteins / Q000502:physiology*; D046912:Multiprotein Complexes / Q000378:metabolism; D010963:Plasmodium falciparum / Q000502:physiology*; D000072417:Protein Domains; D060066:Protein Interaction Maps; D055550:Protein Stability; D021381:Protein Transport; D020543:Proteome / Q000378:metabolism*; D015800:Protozoan Proteins / Q000502:physiology*
chemical_list: D026901:Membrane Transport Proteins; D046912:Multiprotein Complexes; D020543:Proteome; D015800:Protozoan Proteins