title: Novel UDP-GalNAc Derivative Structures Provide Insight into the Donor Specificity of Human Blood Group Glycosyltransferase.
abstract: Two closely related glycosyltransferases are responsible for the final step of the biosynthesis of ABO(H) human blood group A and B antigens. The two enzymes differ by only four amino acid residues, which determine whether the enzymes transfer GalNAc from UDP-GalNAc or Gal from UDP-Gal to the H-antigen acceptor. The enzymes belong to the class of GT-A folded enzymes, grouped as GT6 in the CAZy database, and are characterized by a single domain with a metal dependent retaining reaction mechanism. However, the exact role of the four amino acid residues in the specificity of the enzymes is still unresolved. In this study, we report the first structural information of a dual specificity cis-AB blood group glycosyltransferase in complex with a synthetic UDP-GalNAc derivative. Interestingly, the GalNAc moiety adopts an unusual yet catalytically productive conformation in the binding pocket, which is different from the "tucked under" conformation previously observed for the UDP-Gal donor. In addition, we show that this UDP-GalNAc derivative in complex with the H-antigen acceptor provokes the same unusual binding pocket closure as seen for the corresponding UDP-Gal derivative. Despite this, the two derivatives show vastly different kinetic properties. Our results provide a important structural insight into the donor substrate specificity and utilization in blood group biosynthesis, which can very likely be exploited for the development of new glycosyltransferase inhibitors and probes.
authors.affiliation: From the Department of Chemistry, King's College London, Faculty of Natural & Mathematical Sciences, Britannia House, 7 Trinity Street, London SE1 1DB, United Kingdom.
authors.forename: Gerd K
authors.initials: GK
authors.lastname: Wagner
authors.affiliation: the University of East Anglia, School of Pharmacy, Norwich NR47TJ, England, and.
authors.forename: Thomas
authors.initials: T
authors.lastname: Pesnot
authors.affiliation: the Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-1799, Copenhagen V, Denmark.
authors.forename: Monica M
authors.initials: MM
authors.lastname: Palcic
authors.affiliation: the Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-1799, Copenhagen V, Denmark renj@ssi.dk.
authors.forename: Rene
authors.initials: R
authors.lastname: Jørgensen
mesh_terms: D000017:ABO Blood-Group System / Q000235:genetics / Q000378:metabolism*; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D014539:Uridine Diphosphate Sugars / Q000235:genetics / Q000378:metabolism*
chemical_list: D000017:ABO Blood-Group System; D014539:Uridine Diphosphate Sugars; D016695:Glycosyltransferases
keywords: UDP-GalNAc derivative; crystallography; donor specificity; glycosyltransferase; human; kinetics; nuclear magnetic resonance (NMR)