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dc.contributor.authorDormitzer, Philip R.
dc.contributor.authorGreenberg, Harry B.
dc.contributor.authorHarrison, Stephen C.
dc.date.accessioned2019-10-13T16:04:02Z
dc.date.issued2001
dc.identifier.citationDormitzer, P. R., H. B. Greenberg, and S. C. Harrison. 2001. “Proteolysis of Monomeric Recombinant Rotavirus VP4 Yields an Oligomeric VP5* Core.” Journal of Virology 75 (16): 7339–50. doi:10.1128/JVI.75.16.7339-7350.2001.
dc.identifier.issn0022-538X
dc.identifier.issn1070-6321
dc.identifier.issn1098-5514
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:41542831*
dc.description.abstractRotavirus particles are activated for cell entry by trypsin cleavage of the outer capsid spike protein, VP4, into a hemagglutinin, VP8*, and a membrane penetration protein, VP5*. We have purified rhesus rotavirus VP4, expressed in baculovirus-infected insect cells. Purified VP4 is a soluble, elongated monomer, as determined by analytical ultracentrifugation. Trypsin cleaves purified VP4 at a number of sites that are protected on the virion and yields a heterogeneous group of protease-resistant cores of VP5*. The most abundant tryptic VP5* core is trimmed past the N terminus associated with activation for virus entry into cells. Sequential digestion of purified VP4 with chymotrypsin and trypsin generates homogeneous VP8* and VP5* cores (VP8CT and VP5CT, respectively), which have the authentic trypsin cleavages in the activation region. VP8CT is a soluble monomer composed primarily of P-sheets. VP5CT forms sodium dodecyl sulfate-resistant dimers. These results suggest that trypsinization of rotavirus particles triggers a rearrangement in the VP5* region of VP4 to yield the dimeric spikes observed in icosahedral image reconstructions from electron cryomicroscopy of trypsinized rotavirus virions. The solubility of VP5CT and of trypsinized rotavirus particles suggests that the trypsin-triggered conformational change primes VP4 for a subsequent rearrangement that accomplishes membrane penetration. The domains of VP4 defined by protease analysis contain all mapped neutralizing epitopes, sialic acid binding residues, the heptad repeat region, and the membrane permeabilization region. This biochemical analysis of VP4 provides sequence-specific structural information that complements electron cryomicroscopy data and defines targets and strategies for atomic-resolution structural studies.
dc.language.isoen_US
dc.publisherAmerican Society for Microbiology
dash.licenseLAA
dc.titleProteolysis of Monomeric Recombinant Rotavirus VP4 Yields an Oligomeric VP5* Core
dc.typeJournal Article
dc.description.versionVersion of Record
dc.relation.journalJournal of Virology
dash.depositing.authorHarrison, Stephen::29356e859c6d3698df98e8184352069e::600
dc.date.available2019-10-13T16:04:02Z
dash.workflow.comments1Science Serial ID 62753
dc.identifier.doi10.1128/JVI.75.16.7339-7350.2001
dash.source.volume75;16
dash.source.page7339


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