title: Identification of Plasmodium falciparum RhopH3 protein peptides that specifically bind to erythrocytes and inhibit merozoite invasion.
abstract: The identification of sequences involved in binding to erythrocytes is an important step for understanding the molecular basis of merozoite-erythrocyte interactions that take place during invasion of the Plasmodium falciparum malaria parasite into host cells. Several molecules located in the apical organelles (micronemes, rhoptry, dense granules) of the invasive-stage parasite are essential for erythrocyte recognition, invasion, and establishment of the nascent parasitophorous vacuole. Particularly, it has been demonstrated that rhoptry proteins play an important role in binding to erythrocyte surface receptors, among which is the PfRhopH3 protein, which triggers important immune responses in patients from endemic regions. It has also been reported that anti-RhopH3 antibodies inhibit in vitro invasion of erythrocytes, further supporting its direct involvement in erythrocyte invasion processes. In this study, PfRhopH3 consecutive peptides were synthesized and tested in erythrocyte binding assays for identifying those regions mediating binding to erythrocytes. Fourteen PfRhopH3 peptides presenting high specific binding activity were found, whose bindings were saturable and presented nanomolar dissociation constants. These high-activity binding peptides (HABPs) were characterized by having alpha-helical structural elements, as determined by circular dichroism, and having receptors of a possible sialic acid-dependent and/or glycoprotein-dependent nature, as evidenced in enzyme-treated erythrocyte binding assays and further corroborated by cross-linking assay results. Furthermore, these HABPs inhibited merozoite in vitro invasion of normal erythrocytes at 200 microM by up to 60% and 90%, suggesting that some RhopH3 protein regions are involved in the P. falciparum erythrocyte invasion.
authors.affiliation: Fundación Instituto de Inmunología de Colombia, Bogotá 020304, Colombia.
authors.forename: Carlos Giovanni
authors.initials: CG
authors.lastname: Pinzón
authors.forename: Hernando
authors.initials: H
authors.lastname: Curtidor
authors.forename: Claudia
authors.initials: C
authors.lastname: Reyes
authors.forename: David
authors.initials: D
authors.lastname: Méndez
authors.forename: Manuel Elkin
authors.initials: ME
authors.lastname: Patarroyo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D004912:Erythrocytes / Q000276:immunology* / Q000469:parasitology; D006801:Humans; D017780:Malaria Vaccines / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D052940:Merozoites / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology*; D010963:Plasmodium falciparum / Q000276:immunology*; D011485:Protein Binding; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D014357:Trypsin / Q000737:chemistry
chemical_list: D017780:Malaria Vaccines; D010446:Peptide Fragments; D015800:Protozoan Proteins; C065352:rhoptry associated protein, Plasmodium; D014357:Trypsin