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dc.contributor.authorTiralongo, Joeen_US
dc.contributor.authorAshikov, Angelen_US
dc.contributor.authorRoutier, Françoiseen_US
dc.contributor.authorEckhardt, Matthiasen_US
dc.contributor.authorBakker, Hansen_US
dc.contributor.authorGerardy-Schahn, Ritaen_US
dc.contributor.authorvon Itzstein, Marken_US
dc.date.accessioned2017-04-24T07:56:27Z
dc.date.available2017-04-24T07:56:27Z
dc.date.issued2006en_US
dc.date.modified2009-05-27T08:31:26Z
dc.identifier.issn09596658en_US
dc.identifier.doi10.1093/glycob/cwj029en_AU
dc.identifier.urihttp://hdl.handle.net/10072/13713
dc.description.abstractThe architectural conservation of nucleotide sugar transport proteins (NSTs) enabled the theoretical prediction of putative NSTs in diverse gene databases. In the human genome, 17 NST sequences have been identified but only six have been unequivocally characterized with respect to their transport specificities. Defining transport characteristics of recombinant NSTs has become a major challenge because true zero background systems are widely absent. Production of recombinant NSTs in heterologous systems has developed multifunctionality for some NSTs leading to a novel level of complexity in the field. Assuming that (1) the specificity of NSTs is determined at the primary sequence level and (2) the proteins are autonomously functional units, final definition of the substrate specificity will depend on the use of isolated transport proteins. Herein, we describe the first report of the functional expression of mouse CMP-sialic acid transporter (CST) in Escherichia coli and thus provide significant progress towards the production of transporter proteins in quantities suitable for functional and structural analyses. Recovery of the active NST from inclusion bodies was achieved after solubilization with 8 M urea and stepwise renaturation. After reconstitution into phospholipid vesicles, the recombinant protein demonstrated specific transport for CMP-N-acetylneuraminic acid (CMP-Neu5Ac) with no transport of UDP-sugars. Kinetic studies carried out with CMP-Neu5Ac and established CMP-Neu5Ac antagonist's evaluated natural conformation of the reconstituted protein and clearly demonstrate that the transporter acts as a simple mobile carrier.en_US
dc.description.peerreviewedYesen_US
dc.description.publicationstatusYesen_AU
dc.format.extent254957 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglishen_US
dc.language.isoen_AU
dc.publisherOxford University Pressen_US
dc.publisher.placeUnited Statesen_US
dc.publisher.urihttp://glycob.oxfordjournals.org/en_AU
dc.relation.ispartofstudentpublicationNen_AU
dc.relation.ispartofpagefrom73en_US
dc.relation.ispartofpageto81en_US
dc.relation.ispartofissue1en_US
dc.relation.ispartofjournalGlycobiologyen_US
dc.relation.ispartofvolume16en_US
dc.rights.retentionYen_AU
dc.subject.fieldofresearchcode270301en_US
dc.subject.fieldofresearchcode270199en_US
dc.titleFunctional expression of the CMP-sialic acid transporter in Escherichia coli and its identification as a simple mobile carrieren_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
gro.facultyOffice of the Snr Dep Vice Chancellor, Institute for Glycomicsen_US
gro.rights.copyrightCopyright 2006 authors.This is an open access paper. http://creativecommons.org/licenses/by/3.0/ license that permits unrestricted use, provided that the paper is properly attributed.en_AU
gro.date.issued2006
gro.hasfulltextFull Text


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