Structural requirement of heparan sulfate for interaction with herpes simplex virus type 1 virions and isolated glycoprotein C.
Cell surface heparan sulfates mediate primary attachment of herpes simplex virus type 1, the first step in virus invasion of the cells. Removal of the host cell heparan sulfate results in a significantly diminished susceptibility of the cell to virus infection. On the virus envelope, glycoprotein C has been identified as the major binding site for heparan sulfate in the primary attachment of the virus to host cells. Using selectively desulfated heparins and metabolically labeled host cell heparan sulfate, we have analyzed the structural requirements of heparan sulfate to provide binding sites for glycoprotein C and the whole virus. Employing glycoprotein C affinity chromatography and a virus binding assay, we subfractionated oligosaccharides derived from heparan sulfate and partially desulfated heparin into selectively bound and unbound pools. These were chemically depolymerized and analyzed at the disaccharide level. The shortest glycoprotein C-binding fragment consisted of 10-12 monosaccharide units containing at least one 2-O- and one 6-O-sulfate group that have to be localized in a sequence-specific way, based on the finding that bound and unbound HS fragments do not differ in charge or composition. The binding sequence is found within N-sulfated blocks of heparan sulfate, although several N-acetyl groups can be tolerated within the minimal binding sequence. These minimal requirements for herpes simplex virus type 1 binding to heparan sulfate are clearly distinct from other identified protein binding sites.
Feyzi
E
E
Department of Medical and Physiological Chemistry, Uppsala University, Biomedical Center, S-751 23 Uppsala, Sweden.
Trybala
E
E
Bergström
T
T
Lindahl
U
U
Spillmann
D
D
D000818:Animals; D001665:Binding Sites; D002522:Chlorocebus aethiops; D002999:Clone Cells; D004187:Disaccharides / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism*; D018259:Herpesvirus 1, Human / Q000502:physiology*; D007413:Intestinal Mucosa; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D011485:Protein Binding; D013552:Swine; D014759:Viral Envelope Proteins / Q000302:isolation & purification / Q000378:metabolism*; D014771:Virion / Q000502:physiology
D004187:Disaccharides; D006497:Heparan Sulfate; D009844:Oligosaccharides; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1