OLIGOMERIC STRUCTURE OF GLYCOPROTEINS IN HERPES-SIMPLEX VIRUS TYPE-1

A number of herpes simplex virus (HSV) glycoproteins are found in olig omeric states: glycoprotein E (gE)-gI and gH-gL form heterodimers, and both gB and gC have been detected as homodimers, We have further expl ored the organization of glycoproteins in the virion envelope by using both purified virions to quantitate glycoprotein amounts and proporti ons and chemical cross-linkers to detect oligomers, We purified gB, gC , go, and gH from cells infected with HSV type 1 and used these as imm unological standards, Glycoproteins present in sucrose gradient-purifi ed preparations of two strains of HSV type 1, KOS and NS, were detecte d with antibodies to each of the purified proteins, From these data, g lycoprotein molar ratios of 1:2:11:16 and 1:1:14:9 were calculated for gB/gC/gD/gH in KOS and NS, respectively, gL was also detected in viri ons, although we lacked a purified gL standard for quantitation, We th en asked whether complexes of these glycoproteins could be identified, and if they existed as homo- or hetero-oligomers. Purified KOS was in cubated at 4 degrees C with bis(sulfosuccinimidyl) suberate (BS3), an 11.4 Angstrom (1 Angstrom = 0.1 mm) noncleavable, water-soluble cross- linker, Virus extracts were examined by Western blotting (immunoblotti ng), or immunoprecipitation followed by Western blotting, to assay for homo- and hetero-oligomers. Homodimers of gB, gC, and gD were detecte d, and hetero-oligomers containing gB cross-linked to gC, gC to gD, an d gD to gB were also identified. gH and gL were detected as a hetero-o ligomeric pair and could be cross-linked to gD or gC but not to gB. We conclude that these glycoproteins are capable of forming associations with one another, These studies suggest that glycoproteins are closel y associated in virions and have the potential to function as oligomer ic complexes.