Interaction of the exported malaria protein Pf332 with the red blood cell membrane skeleton.
Intra-erythrocytic Plasmodium falciparum malaria parasites synthesize and export numerous proteins into the red blood cell (RBC) cytosol, where some bind to the RBC membrane skeleton. These interactions are responsible for the altered antigenic, morphological and functional properties of parasite-infected red blood cells (IRBCs). Plasmodium falciparum protein 332 (Pf332) is a large parasite protein that associates with the membrane skeleton and who's function has recently been elucidated. Using recombinant fragments of Pf332 in in vitro interaction assays, we have localised the specific domain within Pf332 that binds to the RBC membrane skeleton to an 86 residue sequence proximal to the C-terminus of Pf332. We have shown that this region partakes in a specific and saturable interaction with actin (K(d)=0.60 microM) but has no detectable affinity for spectrin. The only exported malaria protein previously known to bind to actin is PfEMP3 but here we demonstrate that there is no competition for actin-binding between PfEMP3 and Pf332, suggesting that they bind to different target sequences in actin.
Waller
Karena L
KL
Department of Microbiology, Monash University, VIC 3800, Australia.
Stubberfield
Lisa M
LM
Dubljevic
Valentina
V
Buckingham
Donna W
DW
Mohandas
Narla
N
Coppel
Ross L
RL
Cooke
Brian M
BM
D000199:Actins / Q000378:metabolism; D000818:Animals; D003599:Cytoskeleton / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism / Q000648:ultrastructure; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D010963:Plasmodium falciparum / Q000378:metabolism*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D013049:Spectrin / Q000378:metabolism
D000199:Actins; C411566:EMP3 protein, Plasmodium falciparum; D008565:Membrane Proteins; D015800:Protozoan Proteins; D013049:Spectrin; C073415:antigen Pf332, Plasmodium