Exploiting calnexin expression on phagosomes to isolate Leishmania parasitophorous vacuoles.
We have developed a simple scheme for the isolation of parasitophorous vacuoles (PVs) that harbor Leishmania parasites. This scheme exploits the observation that PVs display endoplasmic reticulum molecules, including the transmembrane protein calnexin. The presence of calnexin at the surface of the PVs distinguishes them from late endosomal vesicles of comparable density. As a result, PVs can be isolated by calnexin affinity selection from an enriched PV fraction obtained by sucrose density fractionation.
Kima
Peter E
PE
Department of Microbiology and Cell Science, University of Florida, Building 981, Box 110700, Gainesville, FL 32611, USA. pkima@ufl.edu
Dunn
Waltraud
W
D000818:Animals; D037281:Calnexin / Q000096:biosynthesis* / Q000737:chemistry; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D007891:Leishmania / Q000302:isolation & purification* / Q000378:metabolism; D008264:Macrophages / Q000469:parasitology* / Q000648:ultrastructure; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D010588:Phagosomes / Q000378:metabolism* / Q000469:parasitology / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000378:metabolism / Q000469:parasitology; D014617:Vacuoles / Q000469:parasitology* / Q000648:ultrastructure
D037281:Calnexin