Mapping of heparin-binding structures on bovine herpesvirus 1 and pseudorabies virus gIII glycoproteins.
The gIII glycoproteins of both bovine herpesvirus 1 (BHV 1) and pseudorabies virus (PrV) mediate the initial and dominant interactions between virus and permissive host cells. By studying virus binding to wild-type and heparin-deficient CHO cells, we demonstrated that the cellular heparin-like moieties play an essential role in BHV 1 and PrV gIII-mediated virus attachment. Subsequent studies were carried out to map the gIII structures that are responsible for heparin binding. First, based on the observation that BHV 1 and PrV are differentially sensitive to heparin inhibition of gIII-mediated attachment to cells, we conducted a gIII domain shuffling experiment. This involved the construction of a set of recombinant BHV 1 expressing BHV 1 and PrV gIII chimeras and then using the sensitivity to heparin inhibition as a means of mapping the potential heparin-binding regions on the gIII molecules. Next, we synthesized panels of partially overlapping BHV 1 and PrV gIII peptides and examined their reactivity to heparin. The results from these experiments demonstrated five heparin-binding sites between amino acid 129 and 310 of BHV 1 gIII and four heparin-binding sites between amino acid 90 and 275 of PrV gIII.
Liang
X
X
Veterinary Infectious Disease Organization, Saskatoon, Saskatchewan, Canada.
Babiuk
L A
LA
Zamb
T J
TJ
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D004252:DNA Mutational Analysis; D004285:Dogs; D000939:Epitopes; D005822:Genetic Vectors / Q000235:genetics; D006493:Heparin / Q000172:deficiency / Q000378:metabolism* / Q000494:pharmacology; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000378:metabolism*; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D000939:Epitopes; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C046998:bovine herpesvirus type-1 glycoproteins; C042966:pseudorabies virus glycoproteins; D006493:Heparin