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dc.contributor.authorMirza, Anne M.
dc.contributor.authorSheehan, John P.
dc.contributor.authorHardy, Larry W.
dc.contributor.authorGlickman, Rhona L.
dc.contributor.authorIorio, Ronald M.
dc.date2022-08-11T08:10:04.000
dc.date.accessioned2022-08-23T16:54:22Z
dc.date.available2022-08-23T16:54:22Z
dc.date.issued1993-10-05
dc.date.submitted2008-08-15
dc.identifier.citationJ Biol Chem. 1993 Oct 5;268(28):21425-31.
dc.identifier.issn0021-9258 (Print)
dc.identifier.pmid8407985
dc.identifier.urihttp://hdl.handle.net/20.500.14038/42484
dc.description.abstractThe hemagglutinin-neuraminidase (HN) glycoprotein of paramyxoviruses is anchored in the virion membrane near its amino terminus, protruding from the virion surface to mediate attachment to cellular receptors. Solubilization of HN spikes can be achieved by treatment of virions with detergent and high salt concentrations. When the solubilized HN protein from the Australia-Victoria (AV) isolate of the virus is incubated at 37 degrees C, a chymotrypsin-sensitive site between residues 112 and 113 is exposed. A chymotrypsin-cleaved soluble form of the protein, named CT-HN, has been prepared using this approach. It is membrane anchor-less, due to removal of a 14-kDa fragment from the NH2 terminus of HN. It retains all potential glycosylation sites and cysteines present in the ectodomain of the native protein. It migrates in nonreducing gels and sediments in sucrose gradients at the rate expected for homodimeric HN. The latter is also consistent with our demonstration by site-directed mutagenesis that cysteine residues at positions 6 and 123, respectively, mediate disulfide-linked homotetramer and homodimer formation. CT-HN retains almost total antigenicity, suggesting that it is conformationally very similar to the intact molecule, as well as receptor recognition function and, at low pH, neuraminidase activity. It should prove to be a useful tool for further studies of the structure and function of this important viral glycoprotein.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=8407985&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.jbc.org/content/268/28/21425.long
dc.subjectAmino Acid Sequence
dc.subjectAnimals
dc.subjectAntibodies, Monoclonal
dc.subjectAntigens, Viral
dc.subjectChick Embryo
dc.subjectChymotrypsin
dc.subjectCysteine
dc.subjectDisulfides
dc.subjectHN Protein
dc.subjectHeat
dc.subjectHydrogen-Ion Concentration
dc.subjectHydrolysis
dc.subjectMembrane Fusion
dc.subjectMolecular Sequence Data
dc.subjectNewcastle disease virus
dc.subjectProtein Conformation
dc.subjectProtein Folding
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleStructure and function of a membrane anchor-less form of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus
dc.typeJournal Article
dc.source.journaltitleThe Journal of biological chemistry
dc.source.volume268
dc.source.issue28
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/829
dc.identifier.contextkey579716
html.description.abstract<p>The hemagglutinin-neuraminidase (HN) glycoprotein of paramyxoviruses is anchored in the virion membrane near its amino terminus, protruding from the virion surface to mediate attachment to cellular receptors. Solubilization of HN spikes can be achieved by treatment of virions with detergent and high salt concentrations. When the solubilized HN protein from the Australia-Victoria (AV) isolate of the virus is incubated at 37 degrees C, a chymotrypsin-sensitive site between residues 112 and 113 is exposed. A chymotrypsin-cleaved soluble form of the protein, named CT-HN, has been prepared using this approach. It is membrane anchor-less, due to removal of a 14-kDa fragment from the NH2 terminus of HN. It retains all potential glycosylation sites and cysteines present in the ectodomain of the native protein. It migrates in nonreducing gels and sediments in sucrose gradients at the rate expected for homodimeric HN. The latter is also consistent with our demonstration by site-directed mutagenesis that cysteine residues at positions 6 and 123, respectively, mediate disulfide-linked homotetramer and homodimer formation. CT-HN retains almost total antigenicity, suggesting that it is conformationally very similar to the intact molecule, as well as receptor recognition function and, at low pH, neuraminidase activity. It should prove to be a useful tool for further studies of the structure and function of this important viral glycoprotein.</p>
dc.identifier.submissionpathoapubs/829
dc.contributor.departmentDepartment of Molecular Genetics and Microbiology
dc.source.pages21425-31


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