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dc.contributor.authorMoustafa, I
dc.contributor.authorConnaris, H
dc.contributor.authorTaylor, M
dc.contributor.authorZaitsev, V
dc.contributor.authorWilson, JC
dc.contributor.authorKiefel, MJ
dc.contributor.authorvon Itzstein, M
dc.contributor.authorTaylor, G
dc.contributor.editorHerbert Tabor
dc.date.accessioned2017-05-03T11:05:38Z
dc.date.available2017-05-03T11:05:38Z
dc.date.issued2004
dc.date.modified2012-02-10T02:00:56Z
dc.identifier.issn0021-9258
dc.identifier.doi10.1074/jbc.M404965200
dc.identifier.urihttp://hdl.handle.net/10072/5050
dc.description.abstractVibrio cholerae neuraminidase (VCNA) plays a significant role in the pathogenesis of cholera by removing sialic acid from higher order gangliosides to unmask GM1, the receptor for cholera toxin. We previously showed that the structure of VCNA is composed of a central ?-propeller catalytic domain flanked by two lectin-like domains, however the nature of the carbohydrates recognised by these lectin domains has remained unknown. We present here structures of the enzyme in complex with two substrates, ?2,3-sialyllactose and ?-2,6-sialyllactose. Both substrate complexes reveal the ?-anomer of N-acetylneuraminic acid (Neu5Ac, NANA) bound to the Nterminal lectin domain, thereby revealing the role of this domain. The large number of interactions suggest a relatively high binding affinity for sialic acid, which was confirmed by calorimetry, which gave a Kd~30?M. Saturation transfer difference (STD) NMR using a non-hydrolysable substrate, Neu5,9Ac2-2-S-(?-2,6)-GlcNAc?1Me, was also used to map the ligand interactions at the VCNA lectin binding site. It is well known that VCNA can hydrolyse both ?-2,3- and ?-2,6-linked sialic acid substrates. In this study using ?-2,3-sialyllactose co-crystallised with VCNA it was revealed that the inhibitor 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (Neu5Ac2en, DANA) was bound at the catalytic site. This observation supports the notion that VCNA can produce its own inhibitor and has been further confirmed by 1H NMR analysis. The discovery of the sialic acid-binding site in the N-lectin-like domain suggests that this might help target VCNA to sialic acid-rich environments, thereby enhancing the catalytic efficiency of the enzyme.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherAmerican Society for Biochemistry and molecular Biology Inc
dc.publisher.placeUSA
dc.publisher.urihttp://www.jbc.org/
dc.relation.ispartofpagefrom40819
dc.relation.ispartofpageto40826
dc.relation.ispartofissue39
dc.relation.ispartofjournalJournal of Biological Chemistry
dc.relation.ispartofvolume279
dc.subject.fieldofresearchChemical sciences
dc.subject.fieldofresearchBiological sciences
dc.subject.fieldofresearchBiomedical and clinical sciences
dc.subject.fieldofresearchHistory, heritage and archaeology
dc.subject.fieldofresearchcode34
dc.subject.fieldofresearchcode31
dc.subject.fieldofresearchcode32
dc.subject.fieldofresearchcode43
dc.titleSialic acid recognition by Vibrio cholerae neuraminidase
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.rights.copyright© 2004 American Society for Biochemistry and Molecular Biology. Please refer to the journal website for access to the definitive, published version.
gro.date.issued2004
gro.hasfulltextNo Full Text
gro.griffith.authorvon Itzstein, Mark
gro.griffith.authorKiefel, Milton


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