The cytoplasmic domain of bovine herpesvirus 1 glycoprotein B is important for maintaining conformation and the high-affinity binding site of gB.

Bovine herpesvirus 1 (BHV-1) glycoprotein B (gB) has been shown to interact with two types of receptor on Madin Darby bovine kidney cells. The first receptor is heparan sulfate proteoglycan, whereas the second high-affinity receptor remains unknown. In order to study the structural requirement for gB's high-affinity binding activity, different forms of the gB ectodomain were expressed and compared with authentic gB. By using chemical cross-linking and sucrose gradient centrifugation, we found that BHV-1 gB was able to form dimers. A region between the cleavage site and the transmembrane anchor region, residues 506 to 763, was found to be required for gB oligomerization. Although the purified gBt and gBtM, two truncated forms of gB, formed oligomers, they did not block the high-affinity cellular receptor, suggesting that oligomerization was not the reason for the loss of the high-affinity binding site on gB. However, an N-terminal juxtamembrane region-located epitope recognized by a monoclonal antibody, designated epitope I, was lost from gBt and gBtM, indicating that both truncated gBs are conformationally changed. Therefore, the structure around this particular region may be required for the existence of the gB high-affinity binding site.