Erythrocyte detergent-resistant membrane proteins: their characterization and selective uptake during malarial infection.
Infection of human erythrocytes by the apicomplexan malaria parasite Plasmodium falciparum results in endovacuolar uptake of 4 host proteins that reside in erythrocyte detergent-resistant membranes (DRMs). Whether this vacuolar transport reflects selective uptake of host DRM proteins remains unknown. A further complication is that DRMs of vastly different protein and cholesterol contents have been isolated from erythrocytes. Here we show that isolated DRMs containing the highest cholesterol-to-protein ratio have low protein mass. Liquid chromatography, mass spectrometry, and antibody-based studies reveal that the major DRM proteins are band 3, flotillin-1 and -2, peroxiredoxin-2, and stomatin. Band 3 and stomatin, which reflect the bulk mass of erythrocyte DRM proteins, and all tested non-DRM proteins are excluded from the vacuolar parasite. In contrast, flotillin-1 and -2 and 8 minor DRM proteins are recruited to the vacuole. These data suggest that DRM association is necessary but not sufficient for vacuolar recruitment and there is active, vacuolar uptake of a subset of host DRM proteins. Finally, the 10 internalized DRM proteins show varied lipid and peptidic anchors indicating that, contrary to the prevailing model of apicomplexan vacuole formation, DRM association, rather than lipid anchors, provides the preferred criteria for protein recruitment to the malarial vacuole.
Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
Sean C
SC
Murphy
Benjamin U
BU
Samuel
Travis
T
Harrison
Kaye D
KD
Speicher
David W
DW
Speicher
Marion E
ME
Reid
Rainer
R
Prohaska
Philip S
PS
Low
Michael J
MJ
Tanner
Narla
N
Mohandas
Kasturi
K
Haldar
D000818:Animals; D001798:Blood Proteins; D015153:Blotting, Western; D002784:Cholesterol / Q000378:metabolism; D002853:Chromatography, Liquid; D003593:Cytoplasm / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000469:parasitology*; D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008055:Lipids / Q000737:chemistry; D008288:Malaria / Q000097:blood* / Q000473:pathology*; D013058:Mass Spectrometry; D021962:Membrane Microdomains; D008565:Membrane Proteins / Q000097:blood; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D010455:Peptides / Q000737:chemistry; D010544:Peroxidases / Q000097:blood; D054464:Peroxiredoxins; D010963:Plasmodium falciparum / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D001798:Blood Proteins; D003902:Detergents; D008055:Lipids; D008565:Membrane Proteins; D010455:Peptides; C036860:STOM protein, human; C106354:flotillins; D002784:Cholesterol; D010544:Peroxidases; D054464:Peroxiredoxins