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by Cheryl-Ann Lobo, Marilis Rodriguez, Marion Reid, and Sara Lustigman

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1 by Cheryl-Ann Lobo, Marilis Rodriguez, Marion Reid, and Sara Lustigman
Glycophorin C is the receptor for the Plasmodium falciparum erythrocyte binding ligand PfEBP-2 (baebl)‏ by Cheryl-Ann Lobo, Marilis Rodriguez, Marion Reid, and Sara Lustigman Blood Volume 101(11): June 1, 2003 ©2003 by American Society of Hematology

2 PfEBP-2 binds to aa 14-22 in exon 2 of GPC
PfEBP-2 binds to aa in exon 2 of GPC. (A) The amino acid sequence of GPC with the epitopes (overlined) corresponding to the monoclonal antibodies: Segment 1, 2B-41: antibody against GPC aa 16-22; Segment 2, 2B-39: antibody against GPC aa 1-6; Segme... PfEBP-2 binds to aa in exon 2 of GPC. (A) The amino acid sequence of GPC with the epitopes (overlined) corresponding to the monoclonal antibodies: Segment 1, 2B-41: antibody against GPC aa 16-22; Segment 2, 2B-39: antibody against GPC aa 1-6; Segment 3, 3-23: antibody against GPC aa 13-20; and Segment 4, 2B42: antibody against GPC aa Exon 1 is represented by plain text; exon 2, bold text; exon 3, italicized text; and exon 4, plain text. The underlined amino acids represent the transmembrane domain. (B-C) mAbs against amino acid residues and of GPC inhibit binding of EBP-2 to red cells. Labeled supernatant proteins of P falciparum were mixed with normal RBCs preincubated with the different antibodies, followed by immunoprecipitations of the soluble extract. The bound PfEBP-2 is shown. Lane 1, antibody against GPC aa 16-22; lane 2, antibody against GPC aa 1-6; lane 3, antibody against GPC aa 13-20; lane 4, antibody against GPC aa 45-50; lane K-1, control mAb against the Kell protein; lane NMS, mouse preimmune serum; lane EBP-2, mouse anti–EBP-2-3. Cheryl-Ann Lobo et al. Blood 2003;101: ©2003 by American Society of Hematology

3 The erythrocyte receptor for PfEBP-2 is GPC
The erythrocyte receptor for PfEBP-2 is GPC. (A) The receptor is neuraminidase- and trypsin-sensitive but chymotrypsin-resistant. 35S-labeled proteins derived from parasite culture supernatants were mixed with red blood cells treated with neuraminidase (N... The erythrocyte receptor for PfEBP-2 is GPC. (A) The receptor is neuraminidase- and trypsin-sensitive but chymotrypsin-resistant. 35S-labeled proteins derived from parasite culture supernatants were mixed with red blood cells treated with neuraminidase (N), trypsin (T), or chymotrypsin (CT), or with untreated cells (WT). Separated cells were then lysed and the soluble fraction immunoprecipitated with anti–EBP2-3 antibodies. The immune complex was dissociated in SDS-PAGE buffer and run on 8% gels. The specific bound protein band in the autoradiograph corresponding to PfEBP-2 is shown. (B) PfEBP-2 does not bind to GPC null (Leach phenotype) cells. 35S-labeled proteins were mixed with rare, variant red blood cells lacking GPB (S-s-), GPA [En(a-)], Kell (K0), GPC (Leach), Dombrock [Gy(a-)], or control wild-type (WT) RBCs. The bound parasite ligand was immunoprecipitated with anti–EBP-2-3 antibodies. (C) PfEBP-2 binds differentially to GPC variant cells. Labeled proteins were mixed with identical amounts of Leach, Gerbich (Ge), Yus, or control (WT) RBCs. The bound parasite ligand was immunoprecipitated with anti–EBP-2-3 antibodies. Cheryl-Ann Lobo et al. Blood 2003;101: ©2003 by American Society of Hematology


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