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dc.contributor.advisorField, Rob
dc.contributor.authorSmith, Shona L.
dc.coverage.spatial125 p.en_US
dc.date.accessioned2018-06-19T08:11:34Z
dc.date.available2018-06-19T08:11:34Z
dc.date.issued1997
dc.identifier.urihttp://hdl.handle.net/10023/14221
dc.description.abstractThe chemical synthesis of sulfate and phosphate derivatives of galactose-α-1,4-N-acetylglucosamine-OR (octyl N-acetyllactosamine) and galactose-α-1,3-N- acetylglucosamine-OR [where R= -(CH2)7CH3] are reported here using N-acetylglucosamine and galactose as starting materials. Sialylation of octyl N- acetyllactosamine derivatives was achieved using trans-sialidase. These compounds were evaluated as potential acceptor substrates for five recombinant α-1,3- fucosyltransferases (α-1,3-FucT) and a semi-pure α-1,3/4-FucT (human milk). The kinetic data showed a wide range of acceptor specificities between different recombinant enzymes. Octyl N-acetyllactosamine 6-O-sulfate proved to be an excellent substrate for α-1,3-FucT VI, with a KM of 0.85 ?M. This substrate has a lower KM than any reported substrate for any α-1,3-FucT. An unusual result was observed for octyl N- acetyllactosamine derivatives containing a sulfate or phosphate group at the site of glycosylation. These compounds were found to be good acceptor substrates for α-1,3-FucT VI and milk α-1,3/4-FucT with KM and Vmax values similar to those of the parent compound, octyl N-acetyllactosamine. Preliminary studies show that the product of such a reaction could contain a sulfate or phosphate diester linkage between fucose and octyl LacNAc. If the anionic substituent at the site of glycosylation is being fucosylated, current models proposed for a mechanism involving an enzyme active site base mechanism cannot explain this result. An alternative mechanism has been suggested involving Mn2+ coordination to the hydroxyl group of the acceptor substrate being glycosylated. This mechanism can also be used to explain the unusual kinetic results obtained for substrates containing a sulfate or phosphate group at the site of glycosylation.en_US
dc.language.isoenen_US
dc.publisherUniversity of St Andrews
dc.subject.lccQP702.P6S6
dc.subject.lcshPolysaccharidesen
dc.titleSynthesis and biological evaluation of acceptor substrates for alpha-1,3-fucosyltransferaseen_US
dc.typeThesisen_US
dc.type.qualificationlevelDoctoralen_US
dc.type.qualificationnamePhD Doctor of Philosophyen_US
dc.publisher.institutionThe University of St Andrewsen_US


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