FIG. 2.

Heparin binding capacity of gBpK⁻ mutant glycoprotein. Vero cells were transfected with plasmids encoding wild-type gB (A) and gBpK⁻ (B). Twenty-four hours posttransfection, the cell monolayers were infected with KΔ4BX virus in the presence of [³⁵S]methionine/cysteine, as described in Materials and Methods. Ten hours p.i., the monolayers were harvested and solubilized with 0.1% Triton X-100-containing buffer and equivalent amounts of detergent-extracted protein (lanes 1) and were incubated for 2 h at 4°C with heparin-acrylic beads. The unbound proteins (lanes 2) and proteins eluted from the heparin-acrylic beads with detergent buffer (lanes 3) or detergent buffer supplemented with 10 mg of heparin per ml (lanes 4) were immunoprecipitated with a pool of gB-specific MAbs. Each sample was analyzed by SDS-PAGE and autoradiography. The arrows indicate the positions of gB wild-type or gBpK⁻ mutant glycoproteins.