Structural analogs of sialic acid interfere with the binding of erythrocyte binding antigen-175 to glycophorin A, an interaction crucial for erythrocyte invasion by Plasmodium falciparum.
Plasmodium falciparum causes the most virulent form of malaria and remains a major worldwide health problem. The erythrocytic development of P. falciparum relies on parasite invasion of host erythrocytes, a process mediated in part by the interaction of erythrocyte binding antigen 175 (EBA-175) with the erythrocyte receptor glycophorin A (GA). The binding domain of EBA-175 that interacts with glycophorin A is a approximately 330 residues module called F2. Several studies have shown that F2 recognizes both sialic acids and the protein backbone on glycophorin A. Here, we have developed ELISA-based quantitative F2-GA binding assays. We also performed a series of competitive inhibition assays to block the F2-GA interaction using a variety of sialic acid analogs. Our data show that both 2,3-didehydro-2-deoxy-N-acetyl neuraminic acid (DANA) and 3'-N-acetyl neuraminyl-N-acetyl lactosamine are excellent inhibitors of the F2-GA interaction. Moderate levels of inhibition were also observed with monomers or oligomers of N-acetyl neuraminic acid (sialic acid). Furthermore, we show that DANA is able to significantly inhibit the invasion of erythrocytes by P. falciparum. Together, our ELISA-based binding assays and in vitro inhibition of erythrocyte invasion data suggest that small variations in the structures of DANA and related inhibitors can result in even more potent invasion inhibitory activities. Our studies provide a platform for the development of high potency inhibitors of the F2-GA interaction using high throughput drug discovery technologies. Such compounds may form part of inhibitor cocktails, which aim to block invasion of erythrocytes by P. falciparum.
Bharara
Ritica
R
Malaria Research Group, International Centre for Genetic Engineering and Biotechnology, P.O. Box 10504, Aruna Asaf Ali Marg, New Delhi 110067, India.
Singh
Shailja
S
Pattnaik
Priyabrata
P
Chitnis
Chetan E
CE
Sharma
Amit
A
D000818:Animals; D000953:Antigens, Protozoan / Q000378:metabolism*; D019281:Dimerization; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000469:parasitology*; D006021:Glycophorins / Q000378:metabolism*; D006801:Humans; D016778:Malaria, Falciparum; D019158:N-Acetylneuraminic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology*; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000472:pathogenicity*; D015800:Protozoan Proteins / Q000378:metabolism*
D000953:Antigens, Protozoan; D006021:Glycophorins; D015800:Protozoan Proteins; C065668:erythrocyte-binding antigen 175, Plasmodium; C036137:2-deoxy-2,3-dehydro-N-acetylneuraminic acid; D019158:N-Acetylneuraminic Acid