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Glycoconjugate Journal

, Volume 33, Issue 6, pp 985–994 | Cite as

Molecular dissection of placental malaria protein VAR2CSA interaction with a chemo-enzymatically synthesized chondroitin sulfate library

  • Nobuo SugiuraEmail author
  • Thomas Mandel ClausenEmail author
  • Tatsumasa Shioiri
  • Tobias Gustavsson
  • Hideto Watanabe
  • Ali Salanti
Original Article

Abstract

Placental malaria, a serious infection caused by the parasite Plasmodium falciparum, is characterized by the selective accumulation of infected erythrocytes (IEs) in the placentas of the pregnant women. Placental adherence is mediated by the malarial VAR2CSA protein, which interacts with chondroitin sulfate (CS) proteoglycans present in the placental tissue. CS is a linear acidic polysaccharide composed of repeating disaccharide units of d-glucuronic acid and N-acetyl-d-galactosamine that are modified by sulfate groups at different positions. Previous reports have shown that placental-adhering IEs were associated with an unusually low sulfated form of chondroitin sulfate A (CSA) and that a partially sulfated dodecasaccharide is the minimal motif for the interaction. However, the fine molecular structure of this CS chain remains unclear. In this study, we have characterized the CS chain that interacts with a recombinant minimal CS-binding region of VAR2CSA (rVAR2) using a CS library of various defined lengths and sulfate compositions. The CS library was chemo-enzymatically synthesized with bacterial chondroitin polymerase and recombinant CS sulfotransferases. We found that C-4 sulfation of the N-acetyl-d-galactosamine residue is critical for supporting rVAR2 binding, whereas no other sulfate modifications showed effects. Interaction of rVAR2 with CS is highly correlated with the degree of C-4 sulfation and CS chain length. We confirmed that the minimum structure binding to rVAR2 is a tri-sulfated CSA dodecasaccharide, and found that a highly sulfated CSA eicosasaccharide is a more potent inhibitor of rVAR2 binding than the dodecasaccharides. These results suggest that CSA derivatives may potentially serve as targets in therapeutic strategies against placental malaria.

Keywords

Chemo-enzymatic synthesis Chondroitin sulfate Enzyme-linked immunosorbent assay (ELISA) Placental malaria Surface plasmon resonance (SPR) VAR2SA 

Abbreviations

CS

chondroitin sulfate

GlcA

d-glucuronic acid

GalNAc

N-acetyl-d-galactosamine

dp

degree of polymerization (number of monosaccharides)

DS

degree of sulfation per disaccharide unit

D4S

degree of C-4 sulfation of GalNAc per disaccharide unit

IE

infected erythrocyte by malaria parasite

PM

placental malaria

PfEMP1

Plasmodium falciparum erythrocyte membrane protein 1

VAR2CSA

a PfEMP1 protein

rVAR2

recombinant minimal CS-binding region of VAR2CSA

SPR

surface plasmon resonance

ELISA

enzyme-linked immunosorbent assay

Notes

Acknowledgments

We thank Yoshimi Shintoku for expert technical assistance. This study was supported in part by a Supported Program for the Strategic Research Foundation at Private Universities 2011-2015 (S1101027) and Grants-in-Aid for Scientific Research (#23570176 to NS and #25293096 to HW) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan. This study was also supported in part by the European Research Councils (ERC) through the MalOnco program (TMC and AS).

Supplementary material

10719_2016_9685_MOESM1_ESM.pdf (1 mb)
ESM 1 (PDF 1 mb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Nobuo Sugiura
    • 1
    Email author
  • Thomas Mandel Clausen
    • 2
    Email author
  • Tatsumasa Shioiri
    • 1
  • Tobias Gustavsson
    • 2
  • Hideto Watanabe
    • 1
  • Ali Salanti
    • 2
  1. 1.Institute for Molecular Science of MedicineAichi Medical UniversityNagakuteJapan
  2. 2.Centre for Medical Parasitology, Department of Immunology and MicrobiologyUniversity of CopenhagenCopenhagen KDenmark

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