Calnexin acts as a molecular chaperone during the folding of glycoprotein B of human cytomegalovirus.
Human cytomegalovirus glycoprotein B (gB) is synthesized as a 105-kDa nonglycosylated polypeptide and cotranslationally modified by addition of N-linked oligosaccharides to a 160-kDa precursor in the endoplasmic reticulum (ER). It is then transported to the Golgi complex, where it is endoproteolytically cleaved to form the disulfide-linked mature gp55-116 complex. Pulse-chase experiments demonstrate that the 160-kDa gB precursor was transiently associated with calnexin, a membrane-bound chaperone, in the ER. The association was maximal immediately after synthesis, and they dissociated with a half-time of 15 min. Complete inhibition of binding by tunicamycin or castanospermine indicates the importance of N-linked oligosaccharides for it. Nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that during an initial stage in the biogenesis, the 160-kDa gB precursor was first synthesized as a fully reduced form and rapidly converted to an oxidized form, with a half-time of 18 min. Both forms of the gB precursor could bind to calnexin. The kinetics of the conversion from the fully reduced to the oxidized form coincided with that of dissociation of the 160-kDa gB precursor from calnexin, suggesting that the two steps are closely related.
Yamashita
Y
Y
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Nagoya, Japan.
Shimokata
K
K
Mizuno
S
S
Daikoku
T
T
Tsurumi
T
T
Nishiyama
Y
Y
D017485:1-Deoxynojirimycin / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037281:Calnexin; D002460:Cell Line; D003587:Cytomegalovirus / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007212:Indolizines / Q000494:pharmacology; D018832:Molecular Chaperones / Q000378:metabolism*; D010084:Oxidation-Reduction; D011485:Protein Binding; D017510:Protein Folding; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000378:metabolism*; D054836:Indolizidines
D017485:1-Deoxynojirimycin; D002135:Calcium-Binding Proteins; D037281:Calnexin; D007212:Indolizines; D018832:Molecular Chaperones; D011498:Protein Precursors; D014415:Tunicamycin; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus; C037806:castanospermine; D054836:Indolizidines