title: Base-modified donor analogues reveal novel dynamic features of a glycosyltransferase.
abstract: Glycosyltransferases (GTs) are enzymes that are involved, as Nature's "glycosylation reagents," in many fundamental biological processes including cell adhesion and blood group biosynthesis. Although of similar importance to that of other large enzyme families such as protein kinases and proteases, the undisputed potential of GTs for chemical biology and drug discovery has remained largely unrealized to date. This is due, at least in part, to a relative lack of GT inhibitors and tool compounds for structural, mechanistic, and cellular studies. In this study, we have used a novel class of GT donor analogues to obtain new structural and enzymological information for a representative blood group GT. These analogues interfere with the folding of an internal loop and the C terminus, which are essential for catalysis. Our experiments have led to the discovery of an entirely new active site folding mode for this enzyme family, which can be targeted in inhibitor development, similar to the DFG motif in protein kinases. Taken together, our results provide new insights into substrate binding, dynamics, and utilization in this important enzyme family, which can very likely be harnessed for the rational development of new GT inhibitors and probes.
authors.affiliation: From the Department of Microbiology and Infection Control, Statens Serum Institut, DK-2300 Copenhagen S, Denmark,; the Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-1799 Copenhagen V, Denmark,. Electronic address: renj@ssi.dk.
authors.forename: René
authors.initials: R
authors.lastname: Jørgensen
authors.affiliation: the University of East Anglia, School of Pharmacy, Norwich NR47TJ, United Kingdom.
authors.forename: Thomas
authors.initials: T
authors.lastname: Pesnot
authors.affiliation: the Department of Chemistry, University of Alberta, Edmonton, Alberta T6G2G2, Canada, and.
authors.forename: Ho Jun
authors.initials: HJ
authors.lastname: Lee
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 King's College London, School of Biomedical Sciences, Institute of Pharmaceutical Science and Department of Chemistry, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom. Electronic address: gerd.wagner@kcl.ac.uk.
authors.forename: Gerd K
authors.initials: GK
authors.lastname: Wagner
mesh_terms: D000017:ABO Blood-Group System / Q000737:chemistry* / Q000378:metabolism; D020816:Amino Acid Motifs; D002384:Catalysis; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D017350:N-Acetylgalactosaminyltransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014531:Uridine Diphosphate Galactose / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
chemical_list: D000017:ABO Blood-Group System; D004791:Enzyme Inhibitors; D014531:Uridine Diphosphate Galactose; D014536:Uridine Diphosphate N-Acetylgalactosamine; D017350:N-Acetylgalactosaminyltransferases
keywords: Crystallography; Enzyme Inhibitors; Glycosyltransferases; Kinetics; Molecular Dynamics