Chemical crosslinking of glycoproteins on the envelope of herpes simplex virus.

A base-reversible, 13-A homo-bifunctional chemical crosslinking reagent, bis[2-(succinimidooxycarbonyloxy)ethyl]sulfone (BSOCOES), was tested for its ability to crosslink the herpes simplex virus type 1 glycoproteins associated with the envelope of purified virions. The crosslinked proteins were fractionated and analyzed by immunoblotting, immunoprecipitation, and two-dimensional polyacrylamide gel electrophoresis using monospecific antisera to detect the glycoproteins, gB and gC. Each of the glycoproteins exhibited distinct patterns of crosslinking. Monomers of gB were crosslinked and appeared as a high-molecular-weight complex. No apparent intermediates were observed between the gB monomer and the high-molecular-weight complex. Analysis of the components in the gB high-molecular-weight complex suggests that gB represents the major component and exists as a high-molecular-weight oligomer on viral envelopes under native conditions. In contrast, gC was cross-linked at a much lower efficiency and a number of components at intermediate molecular weights were detected. Initial studies on crosslinked products that were detectable with anti-gC antibody suggest that one of these products represents a gC dimer.