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dc.contributor.authorCullen, Lori McGinnes
dc.contributor.authorMorrison, Trudy G.
dc.date2022-08-11T08:09:34.000
dc.date.accessioned2022-08-23T16:36:01Z
dc.date.available2022-08-23T16:36:01Z
dc.date.issued1997-04-01
dc.date.submitted2009-03-26
dc.identifier.citationJ Virol. 1997 Apr;71(4):3083-9.
dc.identifier.issn0022-538X (Print)
dc.identifier.pmid9060670
dc.identifier.urihttp://hdl.handle.net/20.500.14038/38692
dc.description.abstractDeterminants of glycosylation site usage were explored by using the hemagglutinin-neuraminidase (HN) glycoprotein of the paramyxovirus Newcastle disease virus. The amino acid sequence of the HN protein, a type II glycoprotein, has six N-linked glycosylation addition sites, G1 to G6, two of which, G5 and G6, are not used for the addition of carbohydrate (L. McGinnes and T. Morrison, Virology 212:398-410, 1995). The sequence of this protein also has 13 cysteine residues in the ectodomain (C2 to C14). Mutation of either cysteine 13 or cysteine 14 resulted in the addition of another oligosaccharide chain to the protein. These cysteine residues flank the normally unused G6 glycosylation addition site, and mutation of the G6 site eliminated the extra glycosylation found in the cysteine mutants. These results suggested that failure to form an intramolecular disulfide bond resulted in the usage of a normally unused glycosylation site. This conclusion was confirmed by preventing cotranslational disulfide bond formation in cells by using dithiothreitol. Under these conditions, the wild-type protein acquired extra glycosylation, which was eliminated by mutation of the G6 site. These results suggest that localized folding events on the nascent chain, such as disulfide bond formation, which block access to the oligosaccharyl transferase are a determinant of glycosylation site usage.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=9060670&dopt=Abstract">Link to Article in PubMed</a>
dc.subjectAmino Acid Sequence
dc.subjectAnimals
dc.subjectBinding Sites
dc.subjectCOS Cells
dc.subjectCysteine
dc.subjectDisulfides
dc.subjectGlycosylation
dc.subjectHN Protein
dc.subjectMolecular Sequence Data
dc.subjectNewcastle disease virus
dc.subjectProtein Folding
dc.subjectRabbits
dc.subjectRecombinant Fusion Proteins
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleDisulfide bond formation is a determinant of glycosylation site usage in the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus
dc.typeJournal Article
dc.source.journaltitleJournal of virology
dc.source.volume71
dc.source.issue4
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=2546&amp;context=oapubs&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/1547
dc.identifier.contextkey798524
refterms.dateFOA2022-08-23T16:36:01Z
html.description.abstract<p>Determinants of glycosylation site usage were explored by using the hemagglutinin-neuraminidase (HN) glycoprotein of the paramyxovirus Newcastle disease virus. The amino acid sequence of the HN protein, a type II glycoprotein, has six N-linked glycosylation addition sites, G1 to G6, two of which, G5 and G6, are not used for the addition of carbohydrate (L. McGinnes and T. Morrison, Virology 212:398-410, 1995). The sequence of this protein also has 13 cysteine residues in the ectodomain (C2 to C14). Mutation of either cysteine 13 or cysteine 14 resulted in the addition of another oligosaccharide chain to the protein. These cysteine residues flank the normally unused G6 glycosylation addition site, and mutation of the G6 site eliminated the extra glycosylation found in the cysteine mutants. These results suggested that failure to form an intramolecular disulfide bond resulted in the usage of a normally unused glycosylation site. This conclusion was confirmed by preventing cotranslational disulfide bond formation in cells by using dithiothreitol. Under these conditions, the wild-type protein acquired extra glycosylation, which was eliminated by mutation of the G6 site. These results suggest that localized folding events on the nascent chain, such as disulfide bond formation, which block access to the oligosaccharyl transferase are a determinant of glycosylation site usage.</p>
dc.identifier.submissionpathoapubs/1547
dc.contributor.departmentDepartment of Molecular Genetics and Microbiology
dc.source.pages3083-9


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