Cross-linking studies show that herpes simplex virus type 1 glycoprotein C molecules are clustered in the membrane of infected cells.
Chemical cross-linking using ethylene glycol succinimidyl succinate (EGS) and dithiobispropionimidate (DTBP) was performed to determine the association of herpes simplex virus type 1 glycoprotein C (HSV-1 gC) with its nearest neighbours. Human embryonic lung (HEL) cells were infected with HSV-1 strain KOS, treated with EGS, lysed with Nonidet P40, immunoprecipitated with monoclonal antibodies specific for gC, and analysed by SDS-PAGE. These analyses demonstrated the presence of cross-linked complexes that migrated with an apparent Mr in the range 150,000 to 260,000. Two-dimensional SDS-PAGE (non-reduced and then reduced) analyses of HSV-1-infected HEL cells treated with the cleavable cross-linker DTBP demonstrated that molecules that comigrated with gC were the only components of these high Mr complexes. Immunoelectroblot (Western blot) analyses using polyclonal rabbit antiserum specific for gC verified that the high Mr complexes contained gC. These results indicated that gC molecules may be localized in the infected cell membrane as dimers.
Kikuchi
G E
GE
Graduate Program in Cellular and Molecular Biology, University of Michigan Medical School, Ann Arbor 48109-0620.
Glorioso
J C
JC
Nairn
R
R
D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis*; D003432:Cross-Linking Reagents; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D011233:Precipitin Tests; D018139:Simplexvirus; D014759:Viral Envelope Proteins / Q000032:analysis*
D003432:Cross-Linking Reagents; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1