Murine rhonoclonal antibodies (MAbs) were made to the 52000 (gp52) and the 93000 to 130000 Mr (gp93–130) glycoproteins from a human cytomegalovirus (HCMV) glycoprotein complex designated gC-I or the gB homologue. MAbs recognizing either gp52 or gp93–130 could immunoprecipitate unreduced gC-I complexes from non-ionic detergent extracts of HCMV. Western blotting was performed with immunoaffmity- purified gC-I complexes which were reduced prior to analysis. MAbs made against gp52 recognized gp52 and a 158000 Mr glycoprotein (gpl58). MAbs which recognized gp93–130 in a Western blot also reacted with gpl58, which is a gC-I precursor glycoprotein. The origin of gp93–130 was demonstrated by the reactivity of our gp93–130 MAbs with a recombinant protein containing the N-terminal portion of the gB gene. These data are consistent with the hypothesis that gp52 and gp93–130 are generated from the same high Mr precursor by proteolysis. MAbs recognizing either gp52 or gp93–130 neutralized Towne strain HCMV, but MAbs recognizing gp52 required complement to neutralize whereas MAbs recognizing gp93–130 did not. It was also determined that gp93–130 and gpl58 have detectable amounts of 0-linked glycans but gp52 does not, showing a difference in the glycosyla- tion of these glycoproteins. Analysis of gC-I disulphide bonds showed that two types were present, one which was very susceptible to reduction and a second which was less susceptible. These complexes could consist of very susceptible inter-complex disulphide bonds and less susceptible intra-complex disulphide bonds.
BanksT., BinH., KousoulasK., SpaeteR., PachlC., PereiraL.1989; A major neutralizing domain maps within the carboxyl-terminal half of the cleaved cytomegalovirus B glycoprotein. Journal of General Virology 70:979–985
BrittW. J.1984; Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology 135:369–378
BrittW. J., VuglerL., StephensE. B.1988; Induction of complement-dependent and -independent neutralizing antibodies by recombinant-derived human cytomegalovirus gp55-116 (gB). Journal of Virology 62:3309–3318
CranageM. P., KouzaridesT., BankierA. T., SatchwellS., WestonK., TomlinsonP., BarrellB., HartH., BellS. E., MinsonA. C., SmithG. L.1986; Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. EMBO Journal 5:3057–3063
de BruijnF. J., LupskiJ. R.1984; The use of transposon Tn5 mutagenesis in the rapid generation of correlated physical and genetic maps of DNA segments cloned into multicopy plasmids-a review. Gene 27:131–149
GahmbergC. G., HakamoriS.-I.1973; External labeling of cell surface galactose and galactosamine in glycolipid and glycoprotein of human erythrocytes. Journal of Biological Chemistry 248:4311–4317
GretchD. R., SuterM., StinskiM. F.1987; The use of biotinylated monoclonal antibodies and streptavidin affinity chromatography to isolate herpesvirus hydrophobic proteins or glycoproteins. 163:270–277
GretchD. R., KariB., RasmussenL., GehrzR. C., StinskiM. F.1988a; Identification and characterization of three distinct families of glycoprotein complexes in the envelope of human cytomegalovirus. Journal of Virology 62:875–881
GretchD. R., GehrzR. C., StinskiM. F.1988b; Characterization of a human cytomegalovirus glycoprotein complex (gel). Journal of General Virology 69:1205–1215
KariB., GehrzR.1988; Isolation and characterization of a human cytomegalovirus glycoproten containing a high content of O-linked oligosaccharides. Archives of Virology 98:171–188
KariB., LussenhopN., GoertzR., Wabuke-BunotiM., RadekeR., GehrzR.1986; Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes. Journal of Virology 60:345–352
MackettM., SmithG. L., MossB.1987; The construction and characterization of vaccinia virus recombinants expressing foreign genes. In DNA CloningII pp. 191–202GloverD. M. Edited by Washington, D.C: IRL Press;
RasmussenL., MullenaxJ., NelsonR., MeriganT. C.1985; Viral polypeptides detected by a complement dependent neutralizing murine monoclonal antibody to human cytomegalovirus. Journal of Virology 55:274–280
SelimovaL. M., TashenovaA. A., ZaidesV. M.1988; Breaking and rejoining of disulfide bonds in external glycoproteins of influenza virus. Virology 166:591–593
SmileyB. L., LupskiJ. R., SvecP. S., McmackenR., GodsonG. N.1982; Sequence of the Escherichia coli dnaG primase gene and regulation of its expression. Proceedings of the National Academy of Sciences, U.S.A 79:4550–4554
SpaeteR. R., ThayerR. M., ProbertW. S., MasiarzF. R., ChamberlinS. H., RasmussenL., MeriganT. C., PachlC.1988; Human cytomegalovirus strain Towne glycoprotein B is processed by proteolytic cleavage. Virology 167:207–225
TaylorD. L., FellowsL. E., FarrarG. H., NashR. J., Taylor-RobinsonD., MobberleyM. A., RyderT. A., JeffriesD. J., TymsA. S.1988; Loss of infectivity after treatment with castanospermine or related plant alkaloids correlates with aberrant glycoprotein synthesis. Antiviral Research 10:11–26