Metabolic Inhibitors of O-GlcNAc Transferase That Act In Vivo Implicate Decreased O-GlcNAc Levels in Leptin-Mediated Nutrient Sensing

Article abstract of DOI:10.1002/anie.201803254

O-Linked glycosylation of serine and threonine residues of nucleocytoplasmic proteins with N-acetylglucosamine (O-GlcNAc) residues is catalyzed by O-GlcNAc transferase (OGT). O-GlcNAc is conserved within mammals and is implicated in a wide range of physiological processes. Herein, we describe metabolic precursor inhibitors of OGT suitable for use both in cells and in vivo in mice. These 5-thiosugar analogues of N-acetylglucosamine are assimilated through a convergent metabolic pathway, most likely involving N-acetylglucosamine-6-phosphate de-N-acetylase (NAGA), to generate a common OGT inhibitor within cells. We show that of these inhibitors, 2-deoxy-2-N-hexanamide-5-thio-d-glucopyranoside (5SGlcNHex) acts in vivo to induce dose- and time-dependent decreases in O-GlcNAc levels in various tissues. Decreased O-GlcNAc correlates, both in vitro within adipocytes and in vivo within mice, with lower levels of the transcription factor Sp1 and the satiety-inducing hormone leptin, thus revealing a link between decreased O-GlcNAc levels and nutrient sensing in peripheral tissues of mammals.

Full text of DOI:10.1002/anie.201803254

A Journal of the Gesellschaft Deutscher Chemiker
Angewandte
Chemie
International Edition
www.angewandte.org
Accepted Article
Title: Metabolic inhibitors of O-GlcNAc transferase (OGT) that act in
vivo implicate decreased O-GlcNAc levels in leptin-mediated
nutrient sensing.
Authors: Ta-Wei Liu, Wesley F. Zandberg, Tracey M. Gloster, Lehua
Deng, Kelsey D Murray, Xiaoyang Shan, and David Jaro
Vocadlo
This manuscript has been accepted after peer review and appears as an
Accepted Article online prior to editing, proofing, and formal publication
of the final Version of Record (VoR). This work is currently citable by
using the Digital Object Identifier (DOI) given below. The VoR will be
published online in Early View as soon as possible and may be different
to this Accepted Article as a result of editing. Readers should obtain
the VoR from the journal website shown below when it is published
to ensure accuracy of information. The authors are responsible for the
content of this Accepted Article.
To be cited as: Angew. Chem. Int. Ed. 10.1002/anie.201803254
Angew. Chem. 10.1002/ange.201803254
Link to VoR: http://dx.doi.org/10.1002/anie.201803254
http://dx.doi.org/10.1002/ange.201803254

10.1002/anie.201803254
Angewandte Chemie International Edition
COMMUNICATION
Metabolic inhibitors of O-GlcNAc transferase (OGT) that act in vivo implicate
decreased O-GlcNAc levels in leptin-mediated nutrient sensing.1
Tai-Wei Liu,† Wesley F. Zandberg,† Tracey M. Gloster,† Lehua Deng, Kelsey D. Murray, Xiaoyang
Shan and David J. Vocadlo*
Abstract: O-linked glycosylation of serine and threonine residues of
nucleocytoplasmic proteins with N-acetylglucosamine (O-GlcNAc)
residues is catalyzed by O-GlcNAc transferase (OGT). O-GlcNAc is
conserved within mammals and is implicated in a wide range of
physiological processes. Here we describe metabolic precursor
inhibitors of OGT suitable for use both in cells and in vivo in mice.
These 5-thiosugar analogues of N-acetylglucosamine are assimilated
through a convergent metabolic pathway, most likely involving N-
acetylglucosamine-6-phosphate de-N-acetylase (NAGA), to generate
a common OGT inhibitor within cells. Of these inhibitors, we show that
catalytic roles. Unfortunately, similar studies regarding the roles
of decreased O-GlcNAc on mammalian physiology are lacking
because there are no OGT inhibitors suitable for use in vivo.
Given the emerging roles of OGT in nutrient sensing and other
processes, inhibitors of OGT that can be used as research tools
in vivo are of high interest.[5]
One approach to decreasing O-GlcNAc levels using small
molecules has been to use broad-spectrum amidotransferase
inhibitors such as 6-diazo-5-oxo-L-norleucine (DON), which
promiscuously blocks all amidotransferases that biosynthesize
many cellular metabolites including UDP-GlcNAc. High
throughput screening has been pursued to deliver hits that can be
improved upon.[5b, 5d] These leads, however, show modest cellular
activity and limited solubility.[5b] They also exhibit off-target cellular
toxicity.[5b] Another approach[5c] has been to generate a GlcNAc
2-deoxy-2-N-hexanamide-5-thio-D-glucopyranoside
(5SGlcNHex)
acts in vivo to induce dose- and time-dependent decreases in O-
GlcNAc levels in various tissues. Decreased O-GlcNAc correlates,
both in vitro within adipocytes and in vivo within mice, with lower levels
of transcription factor Sp1 and the satiety-inducing hormone leptin –
revealing a link between decreased O-GlcNAc levels and nutrient
sensing in peripheral tissues of mammals.
analogue,
2-acetamido-2-deoxy-5-thio--D-glucopyranose
(5SGlcNAc, 3), which in its per-O-acetylated form (Ac45SGlcNAc,
3-OAc) can diffuse across the plasma membrane. Within cells, 3-
OAc is de-O-acetylated and assimilated via the GlcNAc salvage
pathway (Figure 1) to generate UDP-5SGlcNAc (4), which is a
competitive OGT inhibitor (Ki = 8 μM). Ac45SGlcNAc (3-OAc)
lowers cellular O-GlcNAc levels (EC50 = 0.8 to 5 M). This
metabolic OGT inhibitor is not toxic but suffers from poor aqueous
solubility. Indeed, while often used in cell studies,[6] solubilizing 3-
OAc requires high concentrations of DMSO, making it
incompatible for dosing of mammals.
β-O-linked N-acetylglucosamine (O-GlcNAc) is a dynamic
modification of nucleocytoplasmic proteins that is present in all
multicellular eukaryotes.[1] The attachment of GlcNAc (1, Figure
1) to hydroxyl groups of serine or threonine residues of hundreds
of target proteins is catalyzed by the glycosyltransferase O-
GlcNAc transferase (OGT), which uses uridine diphospho-N-
acetylglucosamine (UDP-GlcNAc, 2) as a donor sugar substrate.
The glycosidase O-GlcNAcase (OGA) cleaves O-GlcNAc off from
proteins.
O-GlcNAc is implicated in various diseases including cancer,
neurodegeneration, cardiovascular disease, and obesity.[2]
Notably, O-GlcNAc levels respond to nutrient availability both
within cells and in vivo in animal models.[1b] Transgenic mice
overexpressing OGT in fat or muscle tissue exhibit elevated
serum leptin and insulin levels in addition to insulin resistance.[3]
Further, deletion of the gene encoding OGT from neurons of the
paraventricular nucleus (PVN) within the hypothalamus of mice
results in uncontrolled eating.[2b]
Notably, OGA inhibitors that are active in vivo have helped
gain insights into the roles of increased O-GlcNAc levels in
mammals. Strikingly, OGA inhibitors sometimes yield different
results from those made using genetic approaches to increase O-
GlcNAc levels[4]; perhaps because OGT and OGA also have non-
[a]
Dr. T.-W. Liu, Dr. W. F. Zandberg,[+] Dr. T. M Gloster,[++ ] Dr. L.
Deng, K. D. Murray, Dr. X. Shan, Prof. D. J. Vocadlo*,
Departments of Chemistry & Molecular Biology and Biochemistry
Simon Fraser University, 8888 University Dr., Burnaby, BC, Canada
E-mail: dvocadlo@sfu.ca
Figure 1. The hexosamine biosynthetic pathway (HBP) and the GlcNAc-
salvage pathway yield UDP-GlcNAc (2). Glutamine fructose amidotransferase
(GFAT) catalyzes the conversion of glucosamine (5) to glucosamine-6-
phosphate (GlcNH2-6PO4; 5-6PO4). Acetylation of 5-6PO4 by glucosamine
acetyltransferase (GAT) yields GlcNAc-6PO4, an intermediate also produced by
GlcNAc kinase (GNK) in the GlcNAc-salvage pathway. The sequential action of
phosphoglucosamine mutase (AGM) and GlcNAc pyrophosphorylase (AGX) on
GlcNAc-6PO4 leads to the formation of UDP-GlcNAc (2). 5SGlcNAc (3) can be
similarly salvaged and converted into UDP-5SGlcNAc (4).
[+] Current address: Department of Chemistry, University of British
Columbia, 3247 University Way, Kelowna, BC, Canada
[
++] Current address: School of Biology, Biomedical Sciences
Research Complex, University of St Andrews, North Haugh, St
Andrews, Fife, UK.
†These autors contributed equally.
Supporting information for this article is given via a link at the end of
the document.
This article is protected by copyright. All rights reserved.

10.1002/anie.201803254
Angewandte Chemie International Edition
COMMUNICATION
Accordingly, to explore the roles of decreased O-GlcNAc levels in
organismal physiology, compounds that act in vivo are of high
interest.
To generate a tool compound for inhibiting OGT in vivo, we
synthesized a panel of water soluble analogues of 5SGlcNAc (3)
possessing various N-acyl substituents (7–17, Figure 2a,b
Scheme S1). We reasoned these hydrophobic N-acyl groups
would confer a balance between hydrophilicity and lipophilicity,
making them water soluble yet able to diffuse into cells.
Furthermore, although the substrate specificity of N-
acetylglucosamine-6-phosphate de-N-acetylase (NAGA) is not
known,[7] we speculated that compounds 7–17, once
phosphorylated, might be substrates for this recently identified
enzyme (Figure 2b). Action of NAGA on phosphorylated 7–17
would lead to formation of a common intermediate, 5-thio-
glucosamine-6-phosphate (5-6PO4), which can be assimilated by
the hexosamine biosynthetic pathway (HBP). However, direct
entry of 7–17 into the GlcNAc-salvage pathway cannot be ruled
out since the specificity of NAGA is unknown and bulkier N-
glycolyl (GlcNGc) and N-azidoacetyl (GlcNAz) containing
analogues enter the HBP.[8]
To measure the potential of 7–17 and their per-O-acetylated
congeners to block OGT activity in cells, we treated cells with
these compounds. Analysis of O-GlcNAc levels of cell lysates
showed only some deprotected analogues were effective in
reducing O-GlcNAc levels (Figure 2c, Figure S1). Of these the
promising 2-hexanamide derivative (5SGlcNHex, 12) showed
dose-dependent decreases in O-GlcNAc comparable to
Ac45SGlcNAc (3-OAc) (Figure S2).
We next tested whether these compounds were directly
activated as UDP-linked analogues or if they were metabolized by
NAGA. We therefore analyzed the pool of nucleotide sugars from
cells treated with compounds 7, 8, and 12 by capillary
electrophoresis (CE) (Figure 3). Electropherograms for nucleotide
sugars from cells treated with 7, 8, and 12 revealed two new
peaks that had CE mobilities matching those of the
chemoenzymatically-prepared standards UDP-5SGlcNAc (4) and
Figure 3. 5SGlcNR analogues are sequentially processed in cells by GlcNAc
kinase (GNK) and, likely, N-acetylglucosamine-6-phosphate-de-N-acetylase
(NAGA) to generate 5SGlcNH2-6PO4 (6-PO4). a) Nucleotide sugar analysis of
cells treated with selected inhibitors indicated that all the 5SGlcNR analogues
tested (3-OAc, 7, 8, and 12) were converted within cells into two new nucleotide
sugars, indistinguishable from synthetically prepared UDP-5SGlcNAc (4) and
UDP-5SGalNAc. b) 5SGlcNR analogues were phosphorylated by GNK in vitro.
c) 5SGlcNR-6-phosphates were not substrates for AGM or AGX but were readily
hydrolyzed in vitro by NAGA to produce 5SGlcNH2-6PO4 (6-PO4).
its epimer UDP-5SGalNAc (Figure 3a). Extracts from cells treated
with per-O-acetylated 5SGlcNH2 (6-OAc, Figure S3) also yielded
the same two unnatural nucleotide sugars in cells. These data
suggest that these 5SGlcNR analogues are processed within
cells to the common intermediate 2-amino-2-deoxy-5-thio-
glucopyranose 6-phosphate (6-PO4) and then assimilated by the
HBP to form UDP-5SGlcNAc (4).
To clarify the metabolic processing of compounds 7, 8, and
12, we used recombinant enzymes of the mammalian HBP
(Figure 1). 7, 8, and 12 were all phosphorylated by GNK (Figure
3b), yet none of the phosphorylated products were substrates for
AGM except for 7. However, the nucleotide sugar produced in
Figure 2. Synthetic analogues of 5SGlcNAc (3) are metabolized within cells,
leading to decreased O-GlcNAc levels in cells. a) Possible metabolism of
5SGlcNR analogues (7 – 17) by NAGA prior to their entry into the HBP. b)
Synthetic analogues (7 – 17) bearing various N-acyl R groups. c) Effects of
analogues of O-GlcNAc levels of HEK293 cells. C = vehicle (PBS alone).
This article is protected by copyright. All rights reserved.

10.1002/anie.201803254
Angewandte Chemie International Edition
COMMUNICATION
vitro from 7 by the combined HBP enzymes had a different
mobility than that of nucleotide sugar UDP-5SGlcNAc (4) that we
detected in cells treated with compound 7 (Figure S4). We
accordingly tested whether NAGA converts the corresponding 5-
thiosugar-6-PO4 derivatives of 7, 8, and 12 into 5SGlcNH2-6-PO4
(6-6PO4). We confirmed this scenario by digestion of 12-PO4 with
recombinant human NAGA. Electropherograms obtained by CE
analysis of the processing of 12-PO4 by NAGA showed it was
converted to 5SGlcNH2-6-PO4 (6-PO4) (Figure 3c). These data
suggest that 5SGlcNR analogues 7-17, including 7, 8, and the
most potent derivative 5SGlcNHex (12), are phosphorylated
within cells by GNK and then deacylated by NAGA, which we
found has remarkable substrate tolerance. The resulting common
intermediate, 5SGlcNH2-6-PO4 (6-PO4), is then assimilated via
the HBP to form UDP-5SGlcNAc (4), which leads to metabolic
inhibition of OGT and decreased O-GlcNAc levels in tissues.
Previous efforts to use Ac45SGlcNAc (3-OAc) in vivo in
mammals failed because of its poor aqueous solubility. We
therefore evaluated if these new water-soluble metabolic OGT
inhibitors could be used in vivo. Accordingly, we dosed mice by
intraperitoneal (IP) delivery with our most cell active compound
12. A concentration-dependent decrease in spleen O-GlcNAc
levels was observed upon treatment with 12, with apparent effects
at even 3 mg kg-1 (Figure S5a). Compound 12 (300 mg kg-1)
decreased spleen O-GlcNAc levels over time with maximal
inhibition by 8 h that was maintained for at least 48 h (Figure S5b).
12 reduced O-GlcNAc levels in the kidneys, lungs, fat, pancreas,
heart, spleen and muscle tissue but not in the blood or brain
(Figure S5c). Mice injected with high doses of 12 (300 mg kg-1),
became lethargic, which is a known consequence of low leptin
levels.[9] A second injection of 300 mg kg-1 on the second day
caused mice to be moribund, and we discontinued treatment. We
therefore dosed mice IP with a lower dose of 12 (50 mg kg-1, n=3)
and found reduced O-GlcNAc levels in various tissues 16 h after
dosing but no effect in brain, liver, pancreas, and kidney (Figure
4a,b and Figure S6a-c). O-GlcNAc levels returned to baseline
levels after 16 h (Figure 4a). Metabolic inhibitor 12 therefore acts
in vivo to reversibly lower O-GlcNAcylation in various tissue types.
Notably, others have shown Ac45SGlcNAc (3-OAc) appears able
to inhibit other glycosyltransferases in cell lines.[5b] We find,
however, by lectin blot assessment that 12 induced no apparent
changes in other forms of protein glycosylation in all tissues tested
(Figure S7), even at a dose of 300 mg kg-1 after 48 h. Notably,
these observations are consistent with a recent report using
parent compound Ac45SGlcNAc (3-OAc) in cells.[10]
Figure 4. Dosing of mice with 50 mg kg-1 5SGlcNHex (12) reduced O-GlcNAc
levels and impaired the secretion of the hormone leptin. a) Skeletal muscle from
mice dosed with 50 mg kg-1 of 12 showed a transient reduction in O-GlcNAc
levels by immunoblot analysis (CTD110.6). b) Leptin levels as measured by
ELISA decreased in mice dosed with 12 to a minimum at 16 h. c) Dosing with
12 decreased O-GlcNAc and Sp1 levels to a minimum at 16 h in fat pad tissue.
d) Compound 12 lowered GlcNAc-induced leptin secretion from 3T3-L1
adipocytes. Results are given as the means ± SEM of three independent
samples (n = 3). Each independent sample was tested in duplicate; symbols
denote statistical significance (*p < 0.05, **p < 0.01, ***p < 0.001 compared to
control. Student’s t-test).
glucose uptake. Notably, transcription factors C/EBP-α, -β, and
Sp1 all regulate leptin production. Compound 12 lowered levels
of these known O-GlcNAcylated proteins in adipocytes (Figure
S9) and in mice (Figure S6d). These collective data are consistent
with OGT acting as a nutrient sensor in a process coupling HBP
flux to leptin levels (Figure S10).
We envision that future studies to detail dosing regimens with
12 will be important to explore the effects of decreased O-GlcNAc
on varied physiological processes. Experiments will also be
needed to detail the mechanistic links between reduced O-
GlcNAc levels due to in vivo OGT inhibition, impaired leptin
secretion, and apparent hyperphagia. However, these data
provide the first direct correlation between decreased O-GlcNAc
levels and impaired leptin production in vivo. Our findings are in
accord with studies showing that reduced O-GlcNAc lowers levels
Curiously, we observed that mice treated with either low or
high doses (50-300 mg kg-1) of 12 exhibited engorged stomachs
that were full of rodent chow after 16 h (Figure S8). Because
lethargy and excessive food consumption (hyperphagia) are seen
in mice deficient in leptin signaling,[11] we measured the serum
leptin concentration of animals treated with 50 mg kg-1 12 (Figure
4b). Their leptin levels dropped transiently to a minimum at 16 h
(Figure 4b), consistent with the engorged stomachs[9,12]. Leptin
levels were also lower in mice treated with a high dose of 12 (300
mg kg-1) (Figure S9). We therefore hypothesized that OGT
inhibition might impair leptin production (Figure S10).
of Sp1 in cells[6f,13] and OGT having a pivitol role in fat tissues[14]
.
Notably, these data also suggest that regulation of leptin by O-
GlcNAc is bidirectional in vivo, since overexpression of OGT[3],
knock out of OGA,[15], and metabolic upregulation of UDP-GlcNAc
levels[16] all increase leptin expression.
In summary, we describe convenient new tools to inhibit OGT
in cells and in vivo. Strikingly, inhibition of OGT with 5SGlcNHex
(12) provides support for the hypothesis that reduced O-GlcNAc
levels signal impaired nutrient supply in mammals. We expect
these observations will stimulate activity in the creation of
To test this hypothesis we used 3T3-L1 adipocytes and found
these cells secreted less leptin when treated with either 12 (Figure
4d) or Ac45SGlcNAc (3-OAc) (Figure S11) without affecting
This article is protected by copyright. All rights reserved.

10.1002/anie.201803254
Angewandte Chemie International Edition
COMMUNICATION
Chen, Chembiochem 2015, 16, 2571-2575; i) P. H. Chen, T. J.
Smith, J. Wu, P. F. Siesser, B. J. Bisnett, F. Khan, M. Hogue, E.
Soderblom, F. Tang, J. R. Marks, M. B. Major, B. M. Swarts, M.
Boyce, J. T. Chi, EMBO J 2017, 36, 2233-2250; j) P. S. Banerjee,
J. Ma, G.W. Hart, Proc. Natl. Acad. Sci., USA 2015, 112, 6050-
6055.
A. K. Bergfeld, O. M. Pearce, S. L. Diaz, R. Lawrence, D. J.
Vocadlo, B. Choudhury, J. D. Esko, A. Varki, J .Biol. Chem. 2012,
287, 28898-28916.
a) M. S. Macauley, J. Chan, W. F. Zandberg, Y. He, G. E.
Whitworth, K. A. Stubbs, S. A. Yuzwa, A. J. Bennet, A. Varki, G. J.
Davies, D. J. Vocadlo, J Biol Chem 2012, 287, 28882-28897; bD.
J. Vocadlo, H. C. Hang, E. J. Kim, J. A. Hanover, C. R. Bertozzi,
Proc. Natl. Acad. Sci., USA 2003, 100, 9116-9121.
M. A. Pelleymounter, M. J. Cullen, M. B. Baker, R. Hecht, D.
Winters, T. Boone, F. Collins, Science 1995, 269, 540-543.
L. M. Andres, I. W. Blong, A. C. Evans, N. G. Rumachik, T.
Yamaguchi, N. D. Pham, P. Thompson, J. J. Kohler, C. R.
Bertozzi, ACS Chem. Biol. 2017, 12, 2030-2039.
Y. Zhang, R. Proenca, M. Maffei, M. Barone, L. Leopold, J. M.
Friedman, Nature 1994, 372, 425-432.
J. L. Halaas, K. S. Gajiwala, M. Maffei, S. L. Cohen, B. T. Chait, D.
Rabinowitz, R. L. Lallone, S. K. Burley, J. M. Friedman, Science
1995, 269, 543-546.
I. Han, J. E. Kudlow, Mol. Cell Biol. 1997, 17, 2550-2558.
H. B. Ruan, M. O. Dietrich, Z. W. Liu, M. R. Zimmer, M. D. Li, J. P.
Singh, K. Zhang, R. Yin, J. Wu, T. L. Horvath, X. Yang, Cell 2014,
159, 306-317.
C. Keembiyehetty, D. C. Love, K. R. Harwood, O. Gavrilova, M. E.
Comly, J. A. Hanover, J. Biol. Chem. 2015, 290, 7097-7113.
a) J. Wang, R. Liu, M. Hawkins, N. Barzilai, L. Rossetti, Nature
1998, 393, 684-688; b) E. E. Wollaston-Hayden, R. B. Harris, B.
Liu, R. Bridger, Y. Xu, L. Wells, Front Endocrinol. 2014, 5, 223.
additional metabolic OGT inhibtiors, as well as metabolic
inhibitors of other glycosyltransferases for use in vivo. Moreover,
we envision that this chemical strategy of metabolic OGT
inhibition will serve as a valuable complement to using genetic
methods in various rodent models to accelerate studies to
understand the in vivo roles of O-GlcNAc in mammals.
[7]
[8]
Experimental Section: Please see supporting information for full
experimental procedures. All experiments carried out on animals were
approved by the university animal care committee of SFU.
[9]
Acknowledgments: K. Buettner and M. Dearden from the SFU
animal care facility are thanked for their expert advice and assistance.
S. Yuzwa is thanked for early studies on leptin levels. This work was
supported by a grant from the Natural Sciences and Engineering
Research Council of Canada (NSERC) and Canadian Glycomics
Network (GlycoNet). D.J.V. is a scholar of the Michael Smith
Foundation of Health Research (MSFHR) and a Tier II Canada
Research Chair in Chemical Glycobiology. T.M.G was supported by a
Sir Henry Wellcome Post-doctoral Fellowship and a Research Career
Development Fellowship from the Wellcome Trust.
[10]
[11]
[12]
[13]
[14]
[15]
[16]
Keywords: glycoprotein • inhibitors • thiosugar • nucleotide-
sugars • leptin
References:
[1]
[2]
a) C. R. Torres, G. W. Hart, J. Biol. Chem. 1984, 259, 3308-3317;
b) G. W. Hart, M. P. Housley, C. Slawson, Nature 2007, 446,
1017-1022.
a) C. M. Ferrer, V. L. Sodi, M. J. Reginato, J Mol Biol 2016, 428,
3282-3294; b) O. Lagerlof, J. E. Slocomb, I. Hong, Y. Aponte, S.
Blackshaw, G. W. Hart, R. L. Huganir, Science 2016, 351, 1293-
1296; c) S. Dassanayaka, S. P. Jones, Pharmacol. Therapeut.
2014, 142, 62-71.
[3]
[4]
D. A. McClain, W. A. Lubas, R. C. Cooksey, M. Hazel, G. J.
Parker, D. C. Love, J. A. Hanover, Proc. Natl. Acad. Sci., USA
2002, 99, 10695-10699.
a) C. F. Teo, E. G. El-Karim, L. Wells, Glycobiology 2016, 26,
1198-1208; b) M. S. Macauley, Y. He, T. M. Gloster, K. A. Stubbs,
G. J. Davies, D. J. Vocadlo, Chem. Biol. 2011, 17, 937-948; c) M.
S. Macauley, X. Shan, S. A. Yuzwa, T. M. Gloster, D. J. Vocadlo,
Chem. Biol. 2011, 17, 949-958; d) P. J. Stivers, L. Harmonay, A.
Hicks, H. Mehmet, M. Morris, G. M. Robinson, P. R. Strack, M. J.
Savage, D. M. Zaller, I. Zwierzynski, P. E. Brandish, PLoS One
2015, 10, e0145151.
[5]
a) J. Jiang, M. B. Lazarus, L. Pasquina, P. Sliz, S. Walker, Nat
Chem. Biol. 2011, 8, 72-77; b) R. F. Ortiz-Meoz, J. Jiang, M. B.
Lazarus, M. Orman, J. Janetzko, C. Fan, D. Y. Duveau, Z. W. Tan,
C. J. Thomas, S. Walker, ACS Chem. Biol. 2015, 10, 1392-1397;
c) T. M. Gloster, W. F. Zandberg, J. E. Heinonen, D. L. Shen, L.
Deng, D. J. Vocadlo, Nat Chem Biol 2011, 7, 174-181; d) B. J.
Gross, B. C. Kraybill, S. Walker, J, J. Am. Chem. Soc. 2005, 127,
14588-14589; e) R. Trapannone, K. Rafie, D. M. van Aalten,
Biochem. Soc. Trans. 2016, 44, 88-93.
[6]
a) S. Olivier-Van Stichelen, L. Drougat, V. Dehennaut, I. El Yazidi-
Belkoura, C. Guinez, A. M. Mir, J. C. Michalski, A. S. Vercoutter-
Edouart, T. Lefebvre, Oncogenesis 2012, 1, e36; b) P. J. Lund, J.
E. Elias, M. M. Davis, J. Immunol. 2016, 197, 3086-3098; c) M.
Swamy, S. Pathak, K. M. Grzes, S. Damerow, L. V. Sinclair, D. M.
van Aalten, D. A. Cantrell, Nat. Immunol. 2016, 17, 712-720; d) F.
Doll, A. Buntz, A. K. Spate, V. F. Schart, A. Timper, W. Schrimpf,
C. R. Hauck, A. Zumbusch, V. Wittmann, Angew. Chem. Int. Ed.
Engl. 2016, 55, 2262-2266; e) C. M. Speakman, T. C. Domke, W.
Wongpaiboonwattana, K. Sanders, M. Mudaliar, D. M. van Aalten,
G. J. Barton, M. P. Stavridis, Stem Cells 2014, 32, 2605-2615; f) Y.
Zhu, T. W. Liu, S. Cecioni, R. Eskandari, W. F. Zandberg, D. J.
Vocadlo, Nat. Chem. Biol. 2015, 11, 319-325; g) Y. Zhu, T. W. Liu,
Z. Madden, S. A. Yuzwa, K. Murray, S. Cecioni, N. Zachara, D. J.
Vocadlo, J. Mol. Cell Biol. 2016, 8, 2-16; h) W. Lin, L. Gao, X.
This article is protected by copyright. All rights reserved.

10.1002/anie.201803254
Angewandte Chemie International Edition
COMMUNICATION
COMMUNICATION
Tai-Wei Liu,† Wesley F.
Zandberg,† Tracey M.
Gloster,† Lehua Deng,
Kelsey D. Murray,
Xiaoyang Shan and David
J Vocadlo*
New metabolic inhibitors
of OGT enable chemical
control of O-GlcNAc in
vivo in mice and link
decreased O-GlcNAc
modification in fat with
leptin-mediated nutrient
sensing in vivo.
Page No. – Page No.
Metabolic inhibitors of
O-GlcNAc transferase
(OGT) that act in vivo
implicate decreased O-
GlcNAc levels in leptin-
mediated nutrient
sensing.
This article is protected by copyright. All rights reserved.