title: Use of self-assembling GFP to determine protein topology and compartmentalisation in the Plasmodium falciparum-infected erythrocyte.
abstract: In recent years, and largely supported by the increasing use of transfection technology, much research attention has been given to protein trafficking in the Plasmodium falciparum infected red blood cell. By expression of fluorescent reporter proteins, much information has been gained on both the signals and mechanisms directing proteins to their correct sub-cellular localisation within the parasite and infected host cell. Generally however, verification of the observed fluorescent phenotype is carried out using more traditional techniques such as co-immunofluorescence, protease protection, and cell fractionation followed by Western blot. Here we apply a self-assembling split GFP (saGFP) system and show that this can be used to determine both membrane topology and compartmentalisation using transfection technology alone. As an example, we verify the topology of an ER membrane protein, hDer1-1, and of an exported parasite Hsp40 co-chaperone, PFE55. Additionally, we can demonstrate that this system has the potential to be applied to analysis of organellar proteins.
authors.affiliation: Department of Parasitology, Faculty of Biology, Philipps University Marburg, Marburg, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Külzer
authors.forename: Wiebke
authors.initials: W
authors.lastname: Petersen
authors.forename: Avni
authors.initials: A
authors.lastname: Baser
authors.forename: Katharina
authors.initials: K
authors.lastname: Mandel
authors.forename: Jude M
authors.initials: JM
authors.lastname: Przyborski
mesh_terms: D004912:Erythrocytes / Q000737:chemistry* / Q000469:parasitology*; D049452:Green Fluorescent Proteins / Q000032:analysis; D008967:Molecular Biology / Q000379:methods; D010274:Parasitology / Q000379:methods; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000254:growth & development*; D021381:Protein Transport; D015800:Protozoan Proteins / Q000032:analysis*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D013194:Staining and Labeling / Q000379:methods*; D014162:Transfection
chemical_list: D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins