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Identification of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) as the rosetting ligand of the malaria parasite P. falciparum.

Chen Q, Barragan A, Fernandez V, Sundström A, Schlichtherle M, Sahlén A, Carlson J, Datta S, Wahlgren M - J. Exp. Med. (1998)

Bottom Line: Rosetting PfEMP1 contains clusters of glycosaminoglycan-binding motifs.A recombinant fusion protein (Duffy binding-like 1-glutathione S transferase; Duffy binding-like-1-GST) was found to adhere directly to normal erythrocytes, disrupt naturally formed rosettes, block rosette reformation, and bind to a heparin-Sepharose matrix.The adhesive interactions could be inhibited with heparan sulfate or enzymes that remove heparan sulfate from the cell surface whereas other enzymes or similar glycosaminoglycans of a like negative charge did not affect the binding.

View Article: PubMed Central - PubMed

Affiliation: Microbiology and Tumor Biology Center, Karolinska Institutet, the Swedish Institute for Infectious Disease Control, S-171 77 Stockholm, Sweden.

ABSTRACT
Severe Plasmodium falciparum malaria is characterized by excessive sequestration of infected and uninfected erythrocytes in the microvasculature of the affected organ. Rosetting, the adhesion of P. falciparum-infected erythrocytes to uninfected erythrocytes is a virulent parasite phenotype associated with the occurrence of severe malaria. Here we report on the identification by single-cell reverse transcriptase PCR and cDNA cloning of the adhesive ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1). Rosetting PfEMP1 contains clusters of glycosaminoglycan-binding motifs. A recombinant fusion protein (Duffy binding-like 1-glutathione S transferase; Duffy binding-like-1-GST) was found to adhere directly to normal erythrocytes, disrupt naturally formed rosettes, block rosette reformation, and bind to a heparin-Sepharose matrix. The adhesive interactions could be inhibited with heparan sulfate or enzymes that remove heparan sulfate from the cell surface whereas other enzymes or similar glycosaminoglycans of a like negative charge did not affect the binding. PfEMP1 is suggested to be the rosetting ligand and heparan sulfate, or a heparan sulfate-like molecule, the receptor both for PfEMP1 binding and naturally formed erythrocyte rosettes.

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Map of cDNA structure, sequencing clones, deduced amino acid sequence, and the location of GAG-binding motifs in the rosetting  PfEMP1 of FCR3S1.2. A shows the location of the 434-bp fragment and the three fragments (I, II, and III) that were initially cloned for sequencing. Restriction enzyme digestion sites are indicated by arrows. Additional overlapping clones used for sequencing are shown below. B shows the primary structure of the rosetting FCR3S1.2-PfEMP1. It has two DBL domains (DBL-1 and -4), one CIDR, one TM region, and one ATS. C shows the distribution  of aa in different regions of FCR3S1.2-PfEMP1. D shows the complete aa sequence of FCR3S1.2-PfEMP1. The location of potential GAG-binding  motifs are shown in pink. Motifs No. 4, 5 and 9, 10 (aa 221–232 and 533–549, respectively) are seen as a single stretch since they are located next to each  other. (See Materials and Methods for description of identification of GAG-binding motifs.) These sequence data are available from EMBL/GenBank/ DDBJ under accession number AF003473.
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Figure 2: Map of cDNA structure, sequencing clones, deduced amino acid sequence, and the location of GAG-binding motifs in the rosetting PfEMP1 of FCR3S1.2. A shows the location of the 434-bp fragment and the three fragments (I, II, and III) that were initially cloned for sequencing. Restriction enzyme digestion sites are indicated by arrows. Additional overlapping clones used for sequencing are shown below. B shows the primary structure of the rosetting FCR3S1.2-PfEMP1. It has two DBL domains (DBL-1 and -4), one CIDR, one TM region, and one ATS. C shows the distribution of aa in different regions of FCR3S1.2-PfEMP1. D shows the complete aa sequence of FCR3S1.2-PfEMP1. The location of potential GAG-binding motifs are shown in pink. Motifs No. 4, 5 and 9, 10 (aa 221–232 and 533–549, respectively) are seen as a single stretch since they are located next to each other. (See Materials and Methods for description of identification of GAG-binding motifs.) These sequence data are available from EMBL/GenBank/ DDBJ under accession number AF003473.

Mentions: The entire coding region of the var transcript containing the 434-bp motif was assembled with the 13 overlapping fragments and the coding sequence was found to be composed of 6,684 bp (Fig. 2 D), which encodes a 2,228 amino acid (aa) polypeptide with an estimated molecular weight of 260 kD. A single trypsin-sensitive polypeptide of a similar size was seen after 125I-lactoperoxidase surface labeling of the FCR3S1.2 (Fig. 1 E).


Identification of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) as the rosetting ligand of the malaria parasite P. falciparum.

Chen Q, Barragan A, Fernandez V, Sundström A, Schlichtherle M, Sahlén A, Carlson J, Datta S, Wahlgren M - J. Exp. Med. (1998)

Map of cDNA structure, sequencing clones, deduced amino acid sequence, and the location of GAG-binding motifs in the rosetting  PfEMP1 of FCR3S1.2. A shows the location of the 434-bp fragment and the three fragments (I, II, and III) that were initially cloned for sequencing. Restriction enzyme digestion sites are indicated by arrows. Additional overlapping clones used for sequencing are shown below. B shows the primary structure of the rosetting FCR3S1.2-PfEMP1. It has two DBL domains (DBL-1 and -4), one CIDR, one TM region, and one ATS. C shows the distribution  of aa in different regions of FCR3S1.2-PfEMP1. D shows the complete aa sequence of FCR3S1.2-PfEMP1. The location of potential GAG-binding  motifs are shown in pink. Motifs No. 4, 5 and 9, 10 (aa 221–232 and 533–549, respectively) are seen as a single stretch since they are located next to each  other. (See Materials and Methods for description of identification of GAG-binding motifs.) These sequence data are available from EMBL/GenBank/ DDBJ under accession number AF003473.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199182&req=5

Figure 2: Map of cDNA structure, sequencing clones, deduced amino acid sequence, and the location of GAG-binding motifs in the rosetting PfEMP1 of FCR3S1.2. A shows the location of the 434-bp fragment and the three fragments (I, II, and III) that were initially cloned for sequencing. Restriction enzyme digestion sites are indicated by arrows. Additional overlapping clones used for sequencing are shown below. B shows the primary structure of the rosetting FCR3S1.2-PfEMP1. It has two DBL domains (DBL-1 and -4), one CIDR, one TM region, and one ATS. C shows the distribution of aa in different regions of FCR3S1.2-PfEMP1. D shows the complete aa sequence of FCR3S1.2-PfEMP1. The location of potential GAG-binding motifs are shown in pink. Motifs No. 4, 5 and 9, 10 (aa 221–232 and 533–549, respectively) are seen as a single stretch since they are located next to each other. (See Materials and Methods for description of identification of GAG-binding motifs.) These sequence data are available from EMBL/GenBank/ DDBJ under accession number AF003473.
Mentions: The entire coding region of the var transcript containing the 434-bp motif was assembled with the 13 overlapping fragments and the coding sequence was found to be composed of 6,684 bp (Fig. 2 D), which encodes a 2,228 amino acid (aa) polypeptide with an estimated molecular weight of 260 kD. A single trypsin-sensitive polypeptide of a similar size was seen after 125I-lactoperoxidase surface labeling of the FCR3S1.2 (Fig. 1 E).

Bottom Line: Rosetting PfEMP1 contains clusters of glycosaminoglycan-binding motifs.A recombinant fusion protein (Duffy binding-like 1-glutathione S transferase; Duffy binding-like-1-GST) was found to adhere directly to normal erythrocytes, disrupt naturally formed rosettes, block rosette reformation, and bind to a heparin-Sepharose matrix.The adhesive interactions could be inhibited with heparan sulfate or enzymes that remove heparan sulfate from the cell surface whereas other enzymes or similar glycosaminoglycans of a like negative charge did not affect the binding.

View Article: PubMed Central - PubMed

Affiliation: Microbiology and Tumor Biology Center, Karolinska Institutet, the Swedish Institute for Infectious Disease Control, S-171 77 Stockholm, Sweden.

ABSTRACT
Severe Plasmodium falciparum malaria is characterized by excessive sequestration of infected and uninfected erythrocytes in the microvasculature of the affected organ. Rosetting, the adhesion of P. falciparum-infected erythrocytes to uninfected erythrocytes is a virulent parasite phenotype associated with the occurrence of severe malaria. Here we report on the identification by single-cell reverse transcriptase PCR and cDNA cloning of the adhesive ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1). Rosetting PfEMP1 contains clusters of glycosaminoglycan-binding motifs. A recombinant fusion protein (Duffy binding-like 1-glutathione S transferase; Duffy binding-like-1-GST) was found to adhere directly to normal erythrocytes, disrupt naturally formed rosettes, block rosette reformation, and bind to a heparin-Sepharose matrix. The adhesive interactions could be inhibited with heparan sulfate or enzymes that remove heparan sulfate from the cell surface whereas other enzymes or similar glycosaminoglycans of a like negative charge did not affect the binding. PfEMP1 is suggested to be the rosetting ligand and heparan sulfate, or a heparan sulfate-like molecule, the receptor both for PfEMP1 binding and naturally formed erythrocyte rosettes.

Show MeSH
Related in: MedlinePlus