Characterization of pseudorabies virus glycoprotein C attachment to heparan sulfate proteoglycans.
Pseudorabies virus first attaches to cells through an interaction between the envelope glycoprotein C (gC) and the cell surface heparan sulfate (HS) that is linked to proteoglycans (HSPGs). The HS-binding domain of gC is composed of three discrete heparin-binding domains (HBDs), designated HBD1, -2 and -3 for their proximity to the amino terminus of gC. Each HBD can independently mediate virus attachment to HS, yet each also exhibits a distinct binding preference for differentially sulfated derivatives of heparin. To demonstrate this, affinity columns composed of wild-type gC or mutant gC retaining a single HBD to capture several HSPGs from cultured pig and bovine kidney cells were used. The wild-type gC column bound all of the HSPGs well and, overall, bound more than 90% of the total sample applied to the column. Columns composed of either HBD2 or -3 bound intermediate amounts (40%) of the total sample applied, while the HBD1 column bound low amounts of HSPGs. HBD2 and -3 columns did not uniformly bind all of the HSPGs from bovine kidney cells, but the same HSPGs were bound with equal efficiency on each column. Thus, despite their different preferences for sulfation patterns on HS side-chains, HBD2 and -3 appear to bind the same proteoglycan cores. These results established a hierarchy of HBD2=HBD3>HBD1 in importance for HSPG binding. These in vitro-binding results correlated with the attachment phenotype of virus strains expressing gC with a single HBD in their envelopes.
Rue
Cary A
CA
Department of Molecular Sciences, University of Tennessee Health Science Center, 858 Madison Avenue, Room 201, Memphis, TN 38163, USA1.
Ryan
Patrick
P
Department of Molecular Sciences, University of Tennessee Health Science Center, 858 Madison Avenue, Room 201, Memphis, TN 38163, USA1.
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002482:Cellulose / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011558:Herpesvirus 1, Suid / Q000472:pathogenicity* / Q000502:physiology; D007668:Kidney / Q000166:cytology; D007739:L Cells; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D011509:Proteoglycans / Q000378:metabolism; D011557:Pseudorabies / Q000821:virology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013552:Swine; D053667:Syndecans; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D019812:Heparan Sulfate Proteoglycans; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; D053667:Syndecans; D014759:Viral Envelope Proteins; C042966:pseudorabies virus glycoproteins; D002482:Cellulose