| Autor: |
Bachmann, Anna, Scholz, Judith, JanĂEn, Marthe, Mo-Quen Klinkert, Tannich, Egbert, Bruchhaus, Iris, Petter, Michaela |
| Rok vydání: |
2015 |
| DOI: |
10.6084/m9.figshare.c.3628409_d3.v1 |
| Popis: |
Solubilities of small VSAs analysed by fractionation studies. (A) Overview over selective permeabilization methods. Hypotonic lysis, saponin and streptolysin O differentially permeabilize the membrane system in infected erythrocytes allowing fractionation of distinct subcellular compartments in the supernatant and the pellet. Hypotonic lysis protects contents of the Maurerâ s clefts (MC), while all other proteins released into the supernatant. After saponin treatment soluble contents of the erythrocyte cytoplasm, the MC and the parasitophorous vacuole are released into the supernatant. With streptolysin O, soluble proteins of the erythrocyte cytoplasm are found in the supernatant, while the contents of the parasite, the parasitophorous vacuole and the MC are collected in the pellet. (B and C) Western blot analysis of infected erythrocytes fractions. Trophozoites of the 3D7 parasite strain were enriched by MACS and either lysed by hypotonic lysis (HL), permeabilized with saponin (Sap) or streptolysin O (SLO). The supernatant (SN) and pellet (P) fraction were separated by centrifugation and analysed by western blot analysis. Equivalents of 1x107 cells were loaded in each lane. Blots were probed for the presence of RIFIN, STEVOR and PfMC-2TM using antisera as indicated (B) as well as for the control antigens PfEMP1, SBP1, SERP and PP5 showing correct fractionation of the infected erythrocytes (C). EM: Erythrocyte membrane; HL: Hypotonic lysis; MC: Maurerâ s clefts; P: Pellet fraction; PM: Plasma membrane; PVM Parasitophorous vacuole membrane; Sap: Saponin lysis; SLO: Streptolysin O; SN: Supernatant fraction |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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