A Nonpermeant Biotin Derivative Gains Access to the Parasitophorous Vacuole in Plasmodium falciparum-infected Erythrocytes Permeabilized with Streptolysin O

In its host erythrocyte, the malaria parasite Plasmodium falciparum resides within a parasitophorous vacuole, the membrane of which forms a barrier between the host cell cytosol and the parasite surface. The vacuole is a unique compartment because it contains specific proteins that are believed to b...

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Bibliographic Details
Published in:The Journal of biological chemistry 2002-10, Vol.277 (42), p.40005-40011
Main Authors: Nyalwidhe, Julius, Baumeister, Stefan, Hibbs, Alan R, Tawill, Sallah, Papakrivos, Janni, Volker, Uwe, Lingelbach, Klaus
Format: Article
Language:English
Subjects:
Animals
Bacterial Proteins
Biotin - analogs & derivatives
Biotin - metabolism
Biotin - pharmacology
Biotinylation
Chromatography
Cytosol - metabolism
Electrophoresis, Gel, Two-Dimensional
Erythrocytes - metabolism
Erythrocytes - parasitology
Fluorescent Antibody Technique, Indirect
Humans
Hydrogen-Ion Concentration
Immunoblotting
Plasmodium falciparum - metabolism
Precipitin Tests
Protein Binding
Sepharose - metabolism
Streptavidin - pharmacology
Streptolysins - metabolism
Succinimides - pharmacology
Vacuoles - metabolism
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Summary:In its host erythrocyte, the malaria parasite Plasmodium falciparum resides within a parasitophorous vacuole, the membrane of which forms a barrier between the host cell cytosol and the parasite surface. The vacuole is a unique compartment because it contains specific proteins that are believed to be involved in cell biological functions essential for parasite survival. As a prerequisite for the characterization of the vacuolar proteome, we have developed an experimental approach that allows the selective biotinylation of soluble vacuolar proteins. This approach utilizes nonpermeant biotin derivatives that can be introduced into infected erythrocytes after selective permeabilization of the erythrocyte membrane with the pore-forming protein streptolysin O. The derivatives gain access to the vacuolar lumen but not to the parasite cytosol, thus providing supportive evidence for the existence of nonselective pores within the vacuolar membrane that have been postulated based on electrophysiological studies. Soluble vacuolar proteins that are biotin-labeled can be isolated by affinity chromatography using streptavidin-agarose.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M207077200