Plasmodium falciparum: effects of membrane modulating agents on direct binding of rhoptry proteins to human erythrocytes.
We studied the effects of membrane modulation on the interaction of Plasmodium falciparum rhoptry proteins of 140/130/110 kDa (Rhop-H) with human and mouse erythrocytes. Cells treated with 2-(2-methoxyethoxy)ethyl-8-(cis-2-n-octylcyclopropyl)octanoate, myristoleyl alcohol, and proteins extracted with sublytic concentrations of membrane solubilizing detergents were used in erythrocyte binding assays. Protein binding was evaluated by immunoblotting using Rhop-H- and SERA-specific antisera, 1B9, K15, and 5E3, respectively. Protein binding to liposomes prepared with dipalmitoyl-L-alpha-phosphatidylcholine (DPPC) or dilauroyl-L-alpha-phosphatidylcholine (DLPC) was also examined. Our results show that erythrocyte membrane modulation markedly enhanced direct Rhop-H binding to intact human erythrocytes. Binding of SERA to intact human erythrocytes appeared unaffected. Both DPPC and DLPC liposomes had similar Rhop-H and SERA protein binding activities. However, binding to DLPC liposomes was reduced. Rhop-H and SERA extracted with the detergents octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, sodium deoxycholate, and 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate bound directly to intact human erythrocytes, probably by partitioning hydrophobically into the membranes. Sodium carbonate treatment demonstrated a nonintegral association of Rhop-H with the erythrocyte membrane during invasion. Membrane modulation may expose cryptic phospholipid binding sites in the bilayer.
Ndengele
M M
MM
Department of Biology, Cleveland State University, Ohio 44115, USA.
Messineo
D G
DG
Sam-Yellowe
T
T
Harwalkar
J A
JA
D000818:Animals; D000913:Antibodies, Protozoan / Q000494:pharmacology; D000953:Antigens, Protozoan / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005233:Fatty Alcohols / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D008081:Liposomes / Q000378:metabolism; D051379:Mice; D009439:Neuraminidase / Q000494:pharmacology; D010963:Plasmodium falciparum / Q000276:immunology*; D011485:Protein Binding / Q000187:drug effects; D015800:Protozoan Proteins / Q000378:metabolism*; D013228:Stearates / Q000494:pharmacology; D013347:Subcellular Fractions / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D003902:Detergents; D005233:Fatty Alcohols; D008081:Liposomes; D015800:Protozoan Proteins; D013228:Stearates; C057495:serine repeat antigen, Plasmodium; C018672:9,10-tetradecenol; C023714:A(2)C; D009439:Neuraminidase; D014357:Trypsin