title: Defining the erythrocyte binding domains of Plasmodium vivax tryptophan rich antigen 33.5.
abstract: Tryptophan-rich antigens play important role in host-parasite interaction. One of the Plasmodium vivax tryptophan-rich antigens called PvTRAg33.5 had earlier been shown to be predominantly of alpha helical in nature with multidomain structure, induced immune responses in humans, binds to host erythrocytes, and its sequence is highly conserved in the parasite population. In the present study, we divided this protein into three different parts i.e. N-terminal (amino acid position 24-106), middle (amino acid position 107-192), and C-terminal region (amino acid position 185-275) and determined the erythrocyte binding activity of these fragments. This binding activity was retained by the middle and C-terminal fragments covering 107 to 275 amino acid region of the PvTRAg33.5 protein. Eight non-overlapping peptides covering this 107 to 275 amino acid region were then synthesized and tested for their erythrocyte binding activity to further define the binding domains. Only two peptides, peptide P4 (at 171-191 amino acid position) and peptide P8 (at 255-275 amino acid position), were found to contain the erythrocyte binding activity. Competition assay revealed that each peptide recognizes its own erythrocyte receptor. These two peptides were found to be located on two parallel helices at one end of the protein in the modelled structure and could be exposed on its surface to form a suitable site for protein-protein interaction. Natural antibodies present in the sera of the P. vivax exposed individuals or the polyclonal rabbit antibodies against this protein were able to inhibit the erythrocyte binding activity of PvTRAg33.5, its fragments, and these two synthetic peptides P4 and P8. Further studies on receptor-ligand interaction might lead to the development of the therapeutic reagent.
authors: Department of Biotechnology, All India Institute of Medical Sciences, New Delhi, India.
authors: Hema
authors: H
authors: Bora
authors: Rupesh Kumar
authors: RK
authors: Tyagi
authors: Yagya Dutta
authors: YD
authors: Sharma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004912:Erythrocytes / Q000378:metabolism* / Q000469:parasitology*; D006801:Humans; D016780:Malaria, Vivax / Q000097:blood; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010966:Plasmodium vivax / Q000378:metabolism* / Q000502:physiology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry* / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism; D014364:Tryptophan
chemical_list: D010446:Peptide Fragments; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface; D014364:Tryptophan
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