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How the glycosyltransferase OGT catalyzes amide bond cleavage

Nature Chemical Biology volume 12, pages 899–901 (2016)Cite this article

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Abstract

The essential human enzyme O-linked β-N-acetylglucosamine transferase (OGT), known for modulating the functions of nuclear and cytoplasmic proteins through serine and threonine glycosylation, was unexpectedly implicated in the proteolytic maturation of the cell cycle regulator host cell factor-1 (HCF-1). Here we show that HCF-1 cleavage occurs via glycosylation of a glutamate side chain followed by on-enzyme formation of an internal pyroglutamate, which undergoes spontaneous backbone hydrolysis.

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Figure 1: Two intermediates form during OGT-mediated cleavage of an HCF-1 repeat.
Figure 2: The mechanism of HCF-1 cleavage proceeds via glycosylation on glutamate followed by decomposition to an internal pyroglutamate, which undergoes backbone hydrolysis.

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Acknowledgements

J.J. is a National Science and Engineering Research Council (NSERC) of Canada PGS-M and PGS-D3 fellowship recipient. Funding for this work was provided by a National Institutes of Health grant (R01 GM094263) to S.W. We thank C. Thompson (Bruker Daltonics, Billerica, Massachusetts, USA) for access to and assistance with a SolariX XR 7T q-Q-FT-ICR mass spectrometer. We thank D. Vocadlo (Simon Fraser University, Vancouver, British Columbia, Canada) for providing UDP-5SGlcNAc. X-ray diffraction data were collected at the National Synchrotron Light Source at Brookhaven National Laboratory (Beamline X25).

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Author notes

  1. Michael B Lazarus

    Present address: Present address: Department of Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.,

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    John Janetzko & Michael B Lazarus

  2. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA

    John Janetzko, Michael B Lazarus & Suzanne Walker

  3. Division of Science, Small Molecule Mass Spectrometry, Harvard University, Cambridge, Massachusetts, USA

    Sunia A Trauger

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Contributions

J.J. and S.W. designed research and analyzed data; J.J. performed LC-MS/MS experiments with assistance from S.A.T.; M.B.L. performed X-ray crystallography experiments and solved the structure of hOGT4.5 (D554N); J.J. performed all other experiments reported in the paper; J.J. and S.W. wrote the manuscript with input from all authors.

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Correspondence to Suzanne Walker.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Figures 1–22 and Supplementary Tables 1–11. (PDF 7286 kb)

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Janetzko, J., Trauger, S., Lazarus, M. et al. How the glycosyltransferase OGT catalyzes amide bond cleavage. Nat Chem Biol 12, 899–901 (2016). https://doi.org/10.1038/nchembio.2173

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