The Malaria Circumsporozoite Protein: Interaction of the Conserved Regions I and II-Plus with Heparin-like Oligosaccharides in Heparan Sulfate

Ying, Patrick; Shakibaei, Mehdi; Patankar, Manish S.; Clavijo, Pedro; Beavis, Ronald C.; Clark, Gary F.; Frevert, Ute

doi:10.1006/expr.1996.4134

Cited by 68 publications

(54 citation statements)

References 45 publications

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“…28). In the case of CS, previous work by Ying et al (20) found that CS binds preferentially to more highly sulfated regions of HSPG GAGs. We have extended these studies and found that both CS binding and sporozoite attachment to cells decreases as the degree of GAG chain sulfation decreases.…”

Section: Discussionmentioning

confidence: 94%

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The Binding of the Circumsporozoite Protein to Cell Surface Heparan Sulfate Proteoglycans Is Required for PlasmodiumSporozoite Attachment to Target Cells

Pinzon-Ortiz¹,

Friedman²,

Esko³

et al. 2001

Journal of Biological Chemistry

118

120

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The major surface protein of malaria sporozoites, the circumsporozoite protein, binds to heparan sulfate proteoglycans on the surface of hepatocytes. It has been proposed that this binding event is responsible for the rapid and specific localization of sporozoites to the liver after their injection into the skin by an infected anopheline mosquito. Previous in vitro studies performed under static conditions have failed to demonstrate a significant role for heparan sulfate proteoglycans during sporozoite invasion of cells. We performed sporozoite attachment and invasion assays under more dynamic conditions and found a dramatic decrease in sporozoite attachment to cells in the presence of heparin. In contrast to its effect on attachment, heparin does not appear to have an effect on sporozoite invasion of cells. When substituted heparins were used as competitive inhibitors of sporozoite attachment, we found that sulfation of the glycosaminoglycan chains at both the Nand O-positions was important for sporozoite adhesion to cells. We conclude that the binding of the circumsporozoite protein to hepatic heparan sulfate proteoglycans is likely to function during sporozoite attachment in the liver and that this adhesion event depends on the sulfated glycosaminoglycan chains of the proteoglycans.Protozoans of the genus Plasmodium are the causative agents of malaria. Malaria infection is initiated when an infected anopheline mosquito injects sporozoites during a blood meal. The injected sporozoites travel to the liver and invade hepatocytes where they develop into exoerythrocytic forms. The speed and specificity of sporozoite localization to hepatocytes suggest a receptor-mediated event. Previous studies have shown that the major sporozoite surface protein, the circumsporozoite protein (CS), 1 binds to heparan sulfate proteogly- (3, 4). However, the inhibitory effect on sporozoite infectivity, while demonstrating that the CS-HSPG interaction is important, does not indicate if the glycan is required for sporozoite attachment, invasion, or subsequent development in hepatocytes.In vitro assays (5, 6) have been used to determine whether the CS-HSPG interaction is critical for cell invasion. Frevert et al. (6) found that removal of the majority of cell surface HSPGs had a minimal inhibitory effect on sporozoite invasion of cells. One interpretation of these data is that the binding of CS to HSPGs does not function during sporozoite invasion. Another possibility, however, is that CS binding to HSPGs functions in the more dynamic conditions found in the blood circulation and leads to arrest of sporozoites in the liver sinusoids. In this paper we modify the standard sporozoite invasion assay and provide evidence that the interaction between CS and cell surface HSPGs functions during the initial attachment of sporozoites to cells under conditions that mimic flow. In addition, we show that the sulfate moieties of the HSPG glycosaminoglycan chains (GAGs) are important for attachment of sporozoites to cells. MATERIALS AND METHODSSpo...

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Section: Discussionmentioning

confidence: 94%

“…In addition, CS binds to regions of the GAGs that are more highly sulfated (20,21). In order to determine more precisely if CS binds to the sulfated domains of the GAGs, we performed CS binding assays with HepG2 cells in which the sulfation of the HSPGs was decreased by treating cells with sodium chlorate.…”

Section: Figmentioning

confidence: 99%

The Binding of the Circumsporozoite Protein to Cell Surface Heparan Sulfate Proteoglycans Is Required for PlasmodiumSporozoite Attachment to Target Cells

Pinzon-Ortiz¹,

Friedman²,

Esko³

et al. 2001

Journal of Biological Chemistry

118

120

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“…It is also clear that at least one of the cell surface ligands for CSP is HS (6,29,30), and there have been a number of studies suggesting that region II-plus (region II plus eight downstream residues) serves as the actual HS binding site (21,22,31). However, in these studies, the HS binding activity of region II-plus was determined indirectly in cell culture after HS-lyase digestion by showing diminished binding in Chinese hamster ovary cells deficient in proteoglycan synthesis and by heparin and HS prevention of proteolytic digestion of CSP.…”

Section: Discussionmentioning

confidence: 99%

“…Rathore et al (33) have demonstrated that deletion of region II sequences did not prevent the HS-dependent binding of CSP to hepatocytes. Ying et al (31) found that a synthetic peptide corresponding to region I could inhibit the binding of recombinant CSP to HepG2 cells almost as FIG. 6.…”

Section: Discussionmentioning

confidence: 99%

A Binding Site for Highly Sulfated Heparan Sulfate Is Identified in the N Terminus of the Circumsporozoite Protein

Ancsin

Kisilevsky²

2004

Journal of Biological Chemistry

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Circumsporozoite protein (CSP) coats the malarial sporozoite and functions to target the liver for infection, which is the first step to developing malaria. An important tissue ligand for CSP is the glycosaminoglycan heparan sulfate (HS) found on the surface of hepatocytes and in the basement membrane of the space of Disse. To better understand this efficient targeting process, we set out to identify and characterize the HS binding site(s) of CSP. We synthesized a series of peptides corresponding to five regions of Plasmodium falciparum CSP containing basic residues, a common requirement of HS binding sites, and screened them for heparin and HS binding activity. Only one of these peptides (Pf 2), which contains a motif we have named region I-plus, demonstrated both high affinity heparin/HS binding activity and the ability to block the binding of recombinant CSP to heparin-Sepharose 4B. Analysis by isothermal titration calorimetry revealed that region I-plus has a binding constant of K d ‫؍‬ 5.0 M and a stoichiometry of n ‫؍‬ 7.8 binding sites/heparin chain. Heparin binding was dependent on the amino acid sequence of region I-plus, and the binding sites on heparin/HS are contained within a decasaccharide. Furthermore, HS oligosaccharides rich in sulfate and iduronic acid content (heparin-like) are required for efficient binding. Because liver HS is exceptionally high in both these components relative to the HS of other organs, the HS structural requirements for efficient region I-plus/HS binding are consistent with this peptide sequence functioning to target sporozoites to the liver for attachment to hepatocytes. Finally, the region I-plus heparin/HS binding site was also discovered for two other species that infect humans, Plasmodium malariae and Plasmodium vivax, further supporting the existence of a HS binding domain in the N-terminal portion of CSP.

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“…Region I and region II-plus of CS are known to interact with heparin (6,18,19). All of the mutants bound to the heparin-based affinity column, indicating that the heparin-binding domain in region II-plus doesn't involve cysteine residues or their contribution to the secondary structure.…”

Section: Discussionmentioning

confidence: 99%

Role of cysteines inPlasmodium falciparumcircumsporozoite protein: Interactions with heparin can rejuvenate inactive protein mutants

Rathore

McCutchan

2000

Proc. Natl. Acad. Sci. U.S.A.

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The Malaria Circumsporozoite Protein: Interaction of the Conserved Regions I and II-Plus with Heparin-like Oligosaccharides in Heparan Sulfate

Cited by 68 publications

References 45 publications

The Binding of the Circumsporozoite Protein to Cell Surface Heparan Sulfate Proteoglycans Is Required for PlasmodiumSporozoite Attachment to Target Cells

The Binding of the Circumsporozoite Protein to Cell Surface Heparan Sulfate Proteoglycans Is Required for PlasmodiumSporozoite Attachment to Target Cells

A Binding Site for Highly Sulfated Heparan Sulfate Is Identified in the N Terminus of the Circumsporozoite Protein

Role of cysteines inPlasmodium falciparumcircumsporozoite protein: Interactions with heparin can rejuvenate inactive protein mutants

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