Molecular dissection of placental malaria protein VAR2CSA interaction with a chemo-enzymatically synthesized chondroitin sulfate library.
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.
Institute for Molecular Science of Medicine, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi, 480-1195, Japan. nsugiura@aichi-med-u.ac.jp.
Nobuo
N
Sugiura
Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, CSS, Øster Farigmagsgade 5A, -1014, Copenhagen K, DK, Denmark. tmc@sund.ku.dk.
Thomas Mandel
TM
Clausen
Institute for Molecular Science of Medicine, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi, 480-1195, Japan.
Tatsumasa
T
Shioiri
Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, CSS, Øster Farigmagsgade 5A, -1014, Copenhagen K, DK, Denmark.
Tobias
T
Gustavsson
Institute for Molecular Science of Medicine, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi, 480-1195, Japan.
Hideto
H
Watanabe
Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, CSS, Øster Farigmagsgade 5A, -1014, Copenhagen K, DK, Denmark.
Ali
A
Salanti
D000953:Antigens, Protozoan / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002809:Chondroitin Sulfates / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D016778:Malaria, Falciparum / Q000235:genetics / Q000378:metabolism; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000235:genetics / Q000378:metabolism
D000953:Antigens, Protozoan; C512601:VAR2CSA protein, Plasmodium falciparum; D002809:Chondroitin Sulfates
Chemo-enzymatic synthesis; Chondroitin sulfate; Enzyme-linked immunosorbent assay (ELISA); Placental malaria; Surface plasmon resonance (SPR); VAR2SA