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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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Disruption of preformed, natural rosettes with either DBL-1–GST or ATS–GST.  Results are means and standard  errors of three experiments.
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Figure 4: Disruption of preformed, natural rosettes with either DBL-1–GST or ATS–GST. Results are means and standard errors of three experiments.

Mentions: The effect of the DBL-1 fusion protein on preformed rosettes was studied to confirm the biological role of PfEMP1 in rosetting. Aliquots of a highly rosetting FCR3S1.2 culture were incubated with decreasing concentrations of DBL-1–GST, ATS–GST, or a second DBL-1–GST construct covering a distinct var sequence (var2; reference 6) that lacks GAG-binding motifs. DBL-1 caused a dose-dependent rosette reversion with 40–50% reversion at ∼50 μg/ml, whereas neither the ATS–GST nor the var2 DBL-1 showed any effect on rosettes (Fig. 4 and not shown). The rosettes were not reformed upon prolonged incubation.


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)

Disruption of preformed, natural rosettes with either DBL-1–GST or ATS–GST.  Results are means and standard  errors of three experiments.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 4: Disruption of preformed, natural rosettes with either DBL-1–GST or ATS–GST. Results are means and standard errors of three experiments.
Mentions: The effect of the DBL-1 fusion protein on preformed rosettes was studied to confirm the biological role of PfEMP1 in rosetting. Aliquots of a highly rosetting FCR3S1.2 culture were incubated with decreasing concentrations of DBL-1–GST, ATS–GST, or a second DBL-1–GST construct covering a distinct var sequence (var2; reference 6) that lacks GAG-binding motifs. DBL-1 caused a dose-dependent rosette reversion with 40–50% reversion at ∼50 μg/ml, whereas neither the ATS–GST nor the var2 DBL-1 showed any effect on rosettes (Fig. 4 and not shown). The rosettes were not reformed upon prolonged incubation.

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