Abstract
The 175-kDa erythrocyte binding protein (EBA-175) of Plasmodium falciparum and Duffy antigen binding proteins of P. vivax and P. knowlesi are members of a protein family. The features of this protein family include a cysteine-rich motif present in the erythrocyte receptor-binding domain. We identify here a novel 140-kDa P. falciparum erythrocyte binding protein (EBP2/BAEBL) containing the signature cysteine-rich motif by comparative analysis of gene sequence information. Polyclonal antibodies generated by immunization with an EBP2/BAEBL DNA vaccine immunoprecipitated a 140-kDa protein from P. falciparum schizont-infected erythrocyte lysates. Similar to EBA-175, the binding of EBP2/BAEBL to human erythrocytes was dependent on sialic acids because neuraminidase treatment of those erythrocytes rendered them incapable of binding, but differed from EBA-175 in that trypsin treatment decreased EBP2/BAEBL binding by only twofold compared to a 10-fold reduction in EBA-175 binding. Antibodies raised against the putative erythrocyte-binding domain of EBP2/BAEBL effectively blocked the binding of native EBP2/BAEBL to erythrocytes. These functional antibodies localize EBP2/BAEBL to the invasive apical end of the merozoite. We identify EBP2/BAEBL as a paralogue of EBA-175 and as a novel P. falciparum vaccine candidate.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Motifs
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Amino Acid Sequence
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Animals
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Antigens, Protozoan*
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Blotting, Western
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Carrier Proteins / chemistry
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Carrier Proteins / genetics*
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Carrier Proteins / immunology
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Carrier Proteins / metabolism*
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Cysteine / genetics
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Cysteine / metabolism
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Gene Order
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Humans
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Immunoglobulin G / immunology
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Immunoglobulin G / pharmacology
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Membrane Glycoproteins / metabolism*
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Membrane Proteins
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Plasmodium falciparum / genetics*
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Protein Binding
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Protozoan Proteins / chemistry
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Protozoan Proteins / genetics*
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Protozoan Proteins / immunology
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Protozoan Proteins / metabolism*
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Receptors, Cell Surface / chemistry
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Receptors, Cell Surface / genetics*
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Receptors, Cell Surface / immunology
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Receptors, Cell Surface / metabolism*
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Receptors, Immunologic / metabolism*
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Homology, Amino Acid
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Sialic Acid Binding Ig-like Lectin 1
Substances
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Antigens, Protozoan
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Carrier Proteins
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Duffy antigen binding protein, Plasmodium
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Immunoglobulin G
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Membrane Glycoproteins
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Membrane Proteins
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Protozoan Proteins
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Receptors, Cell Surface
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Receptors, Immunologic
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SIGLEC1 protein, human
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Sialic Acid Binding Ig-like Lectin 1
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Siglec1 protein, mouse
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erythrocyte binding protein-2, Plasmodium falciparum
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erythrocyte-binding antigen 175, Plasmodium
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Cysteine
Associated data
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GENBANK/AF425236
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GENBANK/AY030048
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GENBANK/AY030049
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GENBANK/AY052371