Show simple item record

dc.contributor.authorLamerz, Anne-Christinen_US
dc.contributor.authorHaselhorst, Thomasen_US
dc.contributor.authorvon Itzstein, Marken_US
dc.contributor.authorGerardy-Schahn, Ritaen_US
dc.date.accessioned2017-04-24T07:56:27Z
dc.date.available2017-04-24T07:56:27Z
dc.date.issued2006en_US
dc.date.modified2009-10-19T05:22:55Z
dc.identifier.issn00219258en_US
dc.identifier.doi10.1074/jbc.M600076200en_AU
dc.identifier.urihttp://hdl.handle.net/10072/13714
dc.description.abstractThe dense glycocalyx surrounding the protozoan parasite Leishmania is an essential virulence factor. It protects the parasite from hostile environments in the sandfly vector and mammalian host and supports steps of development and invasion. Therefore, new therapeutic concepts concentrate on disturbing glycocalyx biosynthesis. Deletion of genes involved in the metabolism of galactose and mannose have been shown to drastically reduce Leishmania virulence. Here we report the identification of Leishmania major UDP-glucose pyrophosphorylase (UGP). UGP catalyzes the formation of UDP-glucose from glucose 1-phosphate and UTP. This activation step enables glucose to enter metabolic pathways and is crucial for the activation of galactose. UDP-galactose is made from UDP-glucose by nucleotide-donor transfer to galactose 1-phosphate or by epimerization of the glucose moiety. Isolated in a complementation cloning approach, the activity of L. major UGP was proven in vitro. Moreover, purified protein was used to investigate enzyme kinetics, quaternary organization, and binding of ligands. Whereas sequestration by oligomerization is a known regulatory mechanism for eukaryotic UGPs, the recombinant as well as native L. major UGP migrated as monomer in size exclusion chromatography and in accord with this showed simple Michaelis-Menten kinetics toward all substrates. In saturation transfer difference (STD)-NMR studies, we clearly demonstrated that the molecular geometry at position 4 of glucose is responsible for substrate specificity. Furthermore, the ?-phosphate group of UTP is essential for binding and for induction of the open conformation, which then allows entry of glucose 1-phosphate. Our data provide the first direct proof for the ordered bi-bi mechanism suggested in earlier studies.en_US
dc.description.peerreviewedYesen_US
dc.description.publicationstatusYesen_AU
dc.languageEnglishen_US
dc.language.isoen_AU
dc.publisherThe American Society for Biochemistry and Molecular Biology, Inc.en_US
dc.publisher.placeBethesda, USAen_US
dc.publisher.urihttp://www.jbc.org/en_AU
dc.relation.ispartofstudentpublicationNen_AU
dc.relation.ispartofpagefrom16314en_US
dc.relation.ispartofpageto16322en_US
dc.relation.ispartofissue24en_US
dc.relation.ispartofjournalJournal of Biological Chemistryen_US
dc.relation.ispartofvolume281en_US
dc.rights.retentionYen_AU
dc.subject.fieldofresearchcode270199en_US
dc.subject.fieldofresearchcode249901en_US
dc.subject.fieldofresearchcode270108en_US
dc.titleMolecular Cloning of the Leishmania major UDP-glucose Pyrophosphorylase, Functional Characterization, and Ligand Binding Analyses Using NMR Spectroscopyen_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
gro.date.issued2006
gro.hasfulltextNo Full Text


Files in this item

FilesSizeFormatView

There are no files associated with this item.

This item appears in the following Collection(s)

  • Journal articles
    Contains articles published by Griffith authors in scholarly journals.

Show simple item record