A convenient synthesis of the C-1-phosphonate analogue of UDP-GlcNAc and its evaluation as an inhibitor of O-linked GlcNAc transferase (OGT)

Hajduch, Jan; Nam, Ghilsoo; Kim, Sang Hyun; Fröhlich, Roland; Hanover, John A.; Kirk, Kenneth L.

doi:10.1016/j.carres.2007.10.027

Cited by 48 publications

(40 citation statements)

References 31 publications

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“…1b; Table 1), similar to the previously described weak hOGT inhibitor UDP- C -GlcNAc (Clarke et al 2008; Hajduch et al 2008) (IC 50 = 41 μM, Fig. 1b; Table 1).…”

Section: Resultssupporting

confidence: 84%

Substrate and product analogues as human O-GlcNAc transferase inhibitors

et al. 2010

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Protein glycosylation on serine/threonine residues with N-acetylglucosamine (O-GlcNAc) is a dynamic, inducible and abundant post-translational modification. It is thought to regulate many cellular processes and there are examples of interplay between O-GlcNAc and protein phosphorylation. In metazoa, a single, highly conserved and essential gene encodes the O-GlcNAc transferase (OGT) that transfers GlcNAc onto substrate proteins using UDP–GlcNAc as the sugar donor. Specific inhibitors of human OGT would be useful tools to probe the role of this post-translational modification in regulating processes in the living cell. Here, we describe the synthesis of novel UDP–GlcNAc/UDP analogues and evaluate their inhibitory properties and structural binding modes in vitro alongside alloxan, a previously reported weak OGT inhibitor. While the novel analogues are not active on living cells, they inhibit the enzyme in the micromolar range and together with the structural data provide useful templates for further optimisation.Electronic supplementary materialThe online version of this article (doi:10.1007/s00726-010-0688-y) contains supplementary material, which is available to authorized users.

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“…1b; Table 1), similar to the previously described weak hOGT inhibitor UDP- C -GlcNAc (Clarke et al 2008; Hajduch et al 2008) (IC 50 = 41 μM, Fig. 1b; Table 1).…”

Section: Resultssupporting

confidence: 84%

Substrate and product analogues as human O-GlcNAc transferase inhibitors

et al. 2010

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“…Synthesis of GlcNAc‐1P phosphonate analogue was accomplished starting from the known dimethyl (2,3‐ O ‐isopropylidene‐4,6‐di‐ O ‐benzyl‐α‐ D ‐mannopyranosyl) methanephosphonate (Borodkin et al , 2004) using an adaptation of published methods (Casero et al , 1996; Chang et al , 2006; Hajduch et al , 2008). In essence, after cleavage of the isopropylidene protection, the 3‐OH group was selectively benzoylated through a dibutylstannylene intermediate.…”

Section: Methodsmentioning

confidence: 99%

Structural insights into mechanism and specificity of O-GlcNAc transferase

Clarke

Hurtado‐Guerrero

Pathak

et al. 2008

EMBO J

109

147

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Post-translational modification of protein serines/threonines with N-acetylglucosamine (O-GlcNAc) is dynamic, inducible and abundant, regulating many cellular processes by interfering with protein phosphorylation. O-GlcNAcylation is regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase, both encoded by single, essential, genes in metazoan genomes. It is not understood how OGT recognises its sugar nucleotide donor and performs O-GlcNAc transfer onto proteins/peptides, and how the enzyme recognises specific cellular protein substrates. Here, we show, by X-ray crystallography and mutagenesis, that OGT adopts the (metal-independent) GT-B fold and binds a UDP-GlcNAc analogue at the bottom of a highly conserved putative peptide-binding groove, covered by a mobile loop. Strikingly, the tetratricopeptide repeats (TPRs) tightly interact with the active site to form a continuous 120 Å putative interaction surface, whereas the previously predicted phosphatidylinositide-binding site locates to the opposite end of the catalytic domain. On the basis of the structure, we identify truncation/point mutants of the TPRs that have differential effects on activity towards proteins/peptides, giving first insights into how OGT may recognise its substrates.

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“…40 This phosphonate analog is a poor inhibitor of OGT (IC 50 N 5 mM; the K m for natural substrate is 0.5 μM). 41 The poor inhibitory activity of this compound was attributed to the rigid active-site architecture of OGT, which is able to discriminate between the change in geometry of the O-glycosidic bond (bent geometry) and the change in geometry of the C-glycosidic bond (tetrahedral geometry). However, structural and site-directed mutagenesis studies suggest that the phosphonate analog binds in a similar mode to the natural substrate, 40 although a crystal structure of OGT in a complex with UDP-GlcNAc has not been published to date.…”

Section: Structure Of the Ugm:udp-ch 2 -Galp Complexmentioning

confidence: 99%