Binding of herpes simplex virus to cellular heparan sulphate, an initial step in the adsorption process

Abstract

It has been suggested that heparan sulphate has a receptor function in the initial phase of the attachment of herpes simplex virus (HSV) to cells. We have studied the influence of glycosaminoglycans on cell adsorption of, and plaque formation by, HSV-1 and HSV-2, with regard to the role of saccharide structure, chain length and charge density. Heparin and highly-sulphated heparan sulphate (1-5 sulphate groups/disaccharide unit), but neither chondroitin sulphate nor dermatan sulphate, were found to compete with the cellular receptor for attachment of HSV. Heparan sulphate preparations of low sulphate content (0.5 and 0.7 sulphate groups/disaccharide unit) failed to show any significant interaction with HSV. Oligosaccharides generated by partial deaminative cleavage of heparin were used to determine the minimum molecular size required for the binding of virus; the smallest oligosaccharide which reacted with HSV was composed of l0 monosaccharide units. The importance of charge density was demonstrated more directly by subfractionation of the heparin dodecasaccharide fraction by anion-exchange HPLC. The virus-binding capacities of the four resulting dodecasaccharide subfractions increased from the least sulphated to the most heavily sulphated fraction. The results reported are discussed in relation to virus-receptor interactions involved in the attachment of HSV, including the reported binding of HSV to the fibroblast growth factor receptor.

Structural Requirement of Heparan Sulfate for Interaction with Herpes Simplex Virus Type 1 Virions and Isolated Glycoprotein C

“…The aldehyde groups created by the cleavage were reduced overnight with NaB 3 H 4 followed by incubation with an excess of NaBH 4 for 2-4 h (26). The partially N-desulfated/N-acetylated heparin 12-mer and metabolically labeled HS fragments were cleaved in the same way and reduced with an excess of NaBH 4 . The deamination product of the partially N-desulfated/N-acetylated heparin 12-mer was subjected to gel chromatography on a Bio-Gel P-10 column (1 ϫ 140 cm) in 0.5 M NH 4 HCO 3 .…”

“…Isolated gC-bound and -unbound fractions from 3 H-end-labeled, partially N-desulfated/N-acetylated heparin 12-mers were subjected to deaminative cleavage with nitrous acid, pH 1.5, and the products were reduced with (unlabeled) NaBH 4 . This procedure results in conversion of N-sulfated, but not N-acetylated, GlcN units to 2,5-anhydromannitol (aMan R ) residues, with concomitant cleavage of the corresponding glucosaminidic linkages.…”

“…The first step in viral infection is the attachment to the host cell that enables the virus to penetrate into the cell. Heparan sulfate (HS) on the host cell surface has been implicated in the adherence of numerous microbial agents (2) including human immunodeficiency virus (3), HSV-1 and HSV-2 (4,5). That HS chains act as receptors for HSV-1 was demonstrated by inhibition of attachment to and infection of cells through exogenously added heparin (6) and by the finding that HSV-1 attaches poorly to HS-deficient mutant Chinese hamster ovary cells (5).…”

“…As gC is the major mediator of the virus attachment step, an oligosaccharide capable of binding gC should have an inhibitory effect on HSV-1 infection related to that of native heparin. Earlier in vitro studies have shown that HSV-1 infection can be inhibited by a heparin oligomer consisting of 10 -12 monosaccharide units (4). A more recent study implicated a variety of sulfate groups in the binding of heparin to HSV-1 (13).…”

“…The HS oligomers fractionated with regard to affinity for immobilized gC had been metabolically labeled with [ 3 H]GlcN; deamination products thus were reduced with unlabeled NaBH 4 . The HS oligomers fractionated by interaction with intact HSV-1 virus were metabolically 35 S-labeled; hence, the compositional analysis is restricted to the O-sulfated disaccharides.…”