title: Proteomic and genetic analyses demonstrate that Plasmodium berghei blood stages export a large and diverse repertoire of proteins.
abstract: Malaria parasites actively remodel the infected red blood cell (irbc) by exporting proteins into the host cell cytoplasm. The human parasite Plasmodium falciparum exports particularly large numbers of proteins, including proteins that establish a vesicular network allowing the trafficking of proteins onto the surface of irbcs that are responsible for tissue sequestration. Like P. falciparum, the rodent parasite P. berghei ANKA sequesters via irbc interactions with the host receptor CD36. We have applied proteomic, genomic, and reverse-genetic approaches to identify P. berghei proteins potentially involved in the transport of proteins to the irbc surface. A comparative proteomics analysis of P. berghei non-sequestering and sequestering parasites was used to determine changes in the irbc membrane associated with sequestration. Subsequent tagging experiments identified 13 proteins (Plasmodium export element (PEXEL)-positive as well as PEXEL-negative) that are exported into the irbc cytoplasm and have distinct localization patterns: a dispersed and/or patchy distribution, a punctate vesicle-like pattern in the cytoplasm, or a distinct location at the irbc membrane. Members of the PEXEL-negative BIR and PEXEL-positive Pb-fam-3 show a dispersed localization in the irbc cytoplasm, but not at the irbc surface. Two of the identified exported proteins are transported to the irbc membrane and were named erythrocyte membrane associated proteins. EMAP1 is a member of the PEXEL-negative Pb-fam-1 family, and EMAP2 is a PEXEL-positive protein encoded by a single copy gene; neither protein plays a direct role in sequestration. Our observations clearly indicate that P. berghei traffics a diverse range of proteins to different cellular locations via mechanisms that are analogous to those employed by P. falciparum. This information can be exploited to generate transgenic humanized rodent P. berghei parasites expressing chimeric P. berghei/P. falciparum proteins on the surface of rodent irbc, thereby opening new avenues for in vivo screening adjunct therapies that block sequestration.
authors.affiliation: Biomedical Primate Research Centre, 2288 GJ Rijswijk, The Netherlands.
authors.forename: Erica M
authors.initials: EM
authors.lastname: Pasini
authors.forename: Joanna A
authors.initials: JA
authors.lastname: Braks
authors.forename: Jannik
authors.initials: J
authors.lastname: Fonager
authors.forename: Onny
authors.initials: O
authors.lastname: Klop
authors.forename: Elena
authors.initials: E
authors.lastname: Aime
authors.forename: Roberta
authors.initials: R
authors.lastname: Spaccapelo
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Otto
authors.forename: Matt
authors.initials: M
authors.lastname: Berriman
authors.forename: Jan A
authors.initials: JA
authors.lastname: Hiss
authors.forename: Alan W
authors.initials: AW
authors.lastname: Thomas
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
authors.forename: Chris J
authors.initials: CJ
authors.lastname: Janse
authors.forename: Clemens H M
authors.initials: CH
authors.lastname: Kocken
authors.forename: Blandine
authors.initials: B
authors.lastname: Franke-Fayard
mesh_terms: D000818:Animals; D018955:CD36 Antigens / Q000737:chemistry / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology; D005260:Female; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006790:Host-Parasite Interactions; D008156:Luciferases; D008288:Malaria / Q000378:metabolism* / Q000469:parasitology; D051379:Mice; D009154:Mutation; D010962:Plasmodium berghei / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D021381:Protein Transport; D020543:Proteome / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D052939:Schizonts / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014162:Transfection; D053058:Trophozoites / Q000737:chemistry / Q000378:metabolism*
chemical_list: D018955:CD36 Antigens; D020543:Proteome; D015800:Protozoan Proteins; D049452:Green Fluorescent Proteins; D008156:Luciferases