Addition of high-mannose sugars must precede disulfide bond formation for proper folding of Sendai virus glycoproteins.
The role of glycosylation and of disulfide bonds in the formation of the native structure of the Sendai virus hemagglutinin-neuraminidase (HN) and fusion (F0) glycoproteins was studied. In contrast to the HN and F0 synthesized in vivo, the proteins made from pSP6 transcripts in reticulocyte lysates, whether glycosylated or not, were not recognized by monoclonal antibodies or polyclonal rabbit sera raised against the native proteins; they efficiently reacted only with rabbit antisera raised against the reduced sodium dodecyl sulfate-denatured proteins. These in vitro-made proteins, however, did not contain disulfide bonds. The proteins made in vivo in the presence of tunicamycin, which were also not recognized by the anti-native protein antibodies, did contain disulfide bonds, but they were mainly incorrect interchain disulfide bonds. Moreover, while F0 acquired proper disulfide bonds as soon as it was synthesized under normal conditions in vivo, the disulfides were formed in HN only after a lag of 10 to 30 min. This lag coincides with the delay observed in HN native structure formation. We therefore conclude that the maturation of the HN and F0 proteins depends on the formation of proper intramolecular disulfide bonds, which in turn depends on the previous addition of high-mannose sugars.
Vidal
S
S
Department of Microbiology, University of Geneva Medical School, Switzerland.
Mottet
G
G
Kolakofsky
D
D
Roux
L
L
D003553:Cystine / Q000378:metabolism; D004247:DNA / Q000235:genetics; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D015707:HN Protein; D008358:Mannose / Q000378:metabolism*; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000378:metabolism*
D006023:Glycoproteins; D015707:HN Protein; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D003553:Cystine; D004247:DNA; D008358:Mannose