title: Postoligomerization folding of human cytomegalovirus glycoprotein B: identification of folding intermediates and importance of disulfide bonding.
abstract: Human cytomegalovirus glycoprotein B (gB or UL55) has been demonstrated to be a disulfide-linked homodimer within the envelope of mature virions. Previously, it has been shown that gB undergoes a rapid dimerization nearly coincident with its synthesis. Following dimerization, the molecule slowly folds into a form which can be transported from the endoplasmic reticulum. In this study we have examined the prolonged folding of gB by using a set of defined gB-reactive murine monoclonal antibodies and gB expressed as a recombinant protein in the absence of other human cytomegalovirus proteins. Our results have documented a folding pathway consistent with the relatively rapid dimerization of the translation product followed by delayed conversion into a fully folded molecule. Assembly of the dominant antigenic domain of gB, AD-1, preceded dimerization and folding of the molecule. The fully folded dimer was heat stable, but its conformation was altered by treatment with 2% sodium dodecyl sulfate (SDS), whereas an oligomeric folding intermediate was both heat and SDS stable. Postoligomerization disulfide bond formation could be demonstrated during folding of gB, suggesting that the formation of these covalent bonds could contribute to the prolonged folding of this glycoprotein.
mesh_terms: D001345:Autoradiography; D002460:Cell Line; D003587:Cytomegalovirus / Q000378:metabolism*; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D010084:Oxidation-Reduction; D017510:Protein Folding; D013462:Sulfur Radioisotopes; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D014771:Virion / Q000378:metabolism
chemical_list: D004220:Disulfides; D013462:Sulfur Radioisotopes; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; D008715:Methionine