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dc.contributor.authorHaselhorst, T
dc.contributor.authorWilson, JC
dc.contributor.authorLiakatos, A
dc.contributor.authorKiefel, MJ
dc.contributor.authorDyason, JC
dc.contributor.authorvon Itzstein, M
dc.contributor.editorRonald L. Schnaar
dc.date.accessioned2017-05-03T11:05:38Z
dc.date.available2017-05-03T11:05:38Z
dc.date.issued2004
dc.date.modified2012-02-10T03:01:13Z
dc.identifier.issn0959-6658
dc.identifier.doi10.1093/glycob/cwh108
dc.identifier.urihttp://hdl.handle.net/10072/5048
dc.description.abstractNuclear magnetic resonance (NMR) spectroscopy was used to investigate the transfer of sialic acid from a range of sialic acid donor compounds to acceptor molecules, catalyzed by Trypanosoma cruzi trans-sialidase (TcTS). We demonstrate here that NMR spectroscopy is a powerful tool to monitor the trans-sialidase enzyme reaction for a variety of donor and acceptor molecules. The hydrolysis or transfer reactions that are catalyzed by TcTS were also investigated using a range of N-acetylneuraminosyl-based donor substrates and asialo acceptor molecules. These studies showed that the synthetic N-acetylneuraminosyl donor 4-methylumbelliferyl -D-N-acetylneuraminide (MUN) is hydrolyzed by the enzyme 3-5 times faster than either the disaccharide Neu5Ac(2,3)Gal߱Me or the trisaccharide Neu5Ac(2,3)Lac߱Me. In the transfer reaction, we show that Neu5Ac(2,3)Lac߱Me is the most favorable substrate for TcTS and is a better substrate than the naturally-occurring N-acetylneuraminosyl donor 1-acid glycoprotein. In the case of MUN as the donor molecule, the transfer of Neu5Ac to different acceptors is significantly slower than when other N-acetylneuraminosyl donors are used. We hypothesize that when MUN is bound by the enzyme, the orientation and steric bulk of the umbelliferyl aglycon moiety may restrict the access for the correct positioning of an acceptor molecule. AutoDock studies support our hypothesis and show that the umbelliferyl aglycon moiety undergoes a strong pi-stacking interaction with Trp-312. The binding properties of TcTS towards acceptor (lactose) and donor substrate (Neu5Ac) molecules have also been investigated using saturation transfer difference (STD) NMR experiments. These experiments, taken together with other published data, have clearly demonstrated that lactose in the absence of other coligands does not bind to the TcTS active site or other binding domains. However, in the presence of the sialic acid donor, lactose (an asialo acceptor) was observed by NMR spectroscopy to interact with the enzyme's active site. The association of the asialo acceptor with the active site is an absolute requirement for the transfer reaction to proceed.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.format.extent439878 bytes
dc.format.mimetypeapplication/pdf
dc.languageEnglish
dc.language.isoen_US
dc.publisherOxford University Press
dc.publisher.placeUSA
dc.publisher.urihttp://glycob.oxfordjournals.org/
dc.relation.ispartofpagefrom895
dc.relation.ispartofpageto907
dc.relation.ispartofissue10
dc.relation.ispartofjournalGlycobiology
dc.relation.ispartofvolume14
dc.subject.fieldofresearchBiological Sciences
dc.subject.fieldofresearchMedical and Health Sciences
dc.subject.fieldofresearchcode06
dc.subject.fieldofresearchcode11
dc.titleNMR spectroscopic and molecular modeling investigations of the trans-sialidase from Trypanosoma cruzi.
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
dcterms.licensehttp://creativecommons.org/licenses/by/3.0/
gro.rights.copyright© 2004 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.
gro.date.issued2004
gro.hasfulltextFull Text
gro.griffith.authorvon Itzstein, Mark
gro.griffith.authorWilson, Jenny C.
gro.griffith.authorKiefel, Milton
gro.griffith.authorDyason, Jeffrey C.
gro.griffith.authorHaselhorst, Thomas E.
gro.griffith.authorLiakatos, Angela


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