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Structure of human O-GlcNAc transferase and its complex with a peptide substrate

Nature volume 469, pages 564–567 (2011)Cite this article

Abstract

The essential mammalian enzyme O-linked β-N-acetylglucosamine transferase (O-GlcNAc transferase, here OGT) couples metabolic status to the regulation of a wide variety of cellular signalling pathways by acting as a nutrient sensor1. OGT catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine (UDP-GlcNAc) to serines and threonines of cytoplasmic, nuclear and mitochondrial proteins2,3, including numerous transcription factors4, tumour suppressors, kinases5, phosphatases1 and histone-modifying proteins6. Aberrant glycosylation by OGT has been linked to insulin resistance7, diabetic complications8, cancer9 and neurodegenerative diseases including Alzheimer’s10. Despite the importance of OGT, the details of how it recognizes and glycosylates its protein substrates are largely unknown. We report here two crystal structures of human OGT, as a binary complex with UDP (2.8 Å resolution) and as a ternary complex with UDP and a peptide substrate (1.95 Å). The structures provide clues to the enzyme mechanism, show how OGT recognizes target peptide sequences, and reveal the fold of the unique domain between the two halves of the catalytic region. This information will accelerate the rational design of biological experiments to investigate OGT’s functions; it will also help the design of inhibitors for use as cellular probes and help to assess its potential as a therapeutic target.

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Figure 1: Overall structure of human OGT complexed to UDP.
Figure 2: Structure of the OGT–UDP–peptide complex.
Figure 3: Structure of the intervening domain and full-length models of human OGT.

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Primary accessions

Protein Data Bank

Data deposits

The structures of the OGT–UDP complex and the OGT–UDP–peptide complex have been submitted to the Protein Data Bank under accession numbers 3PE3 and 3PE4. Atomic coordinates for the full-length models of OGT as well as the docked UDP-GlcNAc structure are available for download from the Walker Laboratory website (see Supplementary Information).

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Acknowledgements

We thank B. Gross and C. Drennan for advice. We also thank the US National Institutes of Health, the US National Science Foundation, and the Harvard Biomedical Accelerator Fund for financial support. This work is based on research conducted at the Advanced Photon Source (Northeastern Collaborative Access Team beamlines) and Brookhaven National Laboratory (X25 and X29 beamlines).

Author information

Author notes

  1. Michael B. Lazarus and Yunsun Nam: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, 02138, Massachusetts, USA

    Michael B. Lazarus

  2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Yunsun Nam & Piotr Sliz

  3. Laboratory of Molecular Medicine, Children’s Hospital, Boston, 02115, Massachusetts, USA

    Yunsun Nam & Piotr Sliz

  4. Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Michael B. Lazarus, Jiaoyang Jiang & Suzanne Walker

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Contributions

S.W. conceived the project. M.B.L. obtained the crystallization construct and initial diffracting crystals. M.B.L., Y.N. and P.S. determined and refined the crystal structures. J.J. and M.B.L. performed the enzymatic assays. M.B.L., Y.N., J.J., P.S. and S.W. designed experiments, discussed results, and prepared the manuscript.

Corresponding authors

Correspondence to Piotr Sliz or Suzanne Walker.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-9 with legends, Supplementary Tables 1-6 and Supplementary References. (PDF 1198 kb)

Supplementary Movie 1

This movie shows the molecular dynamics simulations of OGT. It is based on a 1 microsecond simulation and shows the global movement of the TPRs based on motion of the hinge described in Supplementary Figure 3. (MOV 3056 kb)

Supplementary Model 1

PDB coordinates for the model of ncOGT bound to UDP. As described in the caption of Fig. 3c, this full-length model was prepared by combining our OGT-UDP structure (PDB code 3PE3) with the OGT TPR structure (PDB code 1W3B). (TXT 1016 kb)

Supplementary Model 2

PDB coordinates for the model of ncOGT bound to UDP and the CKII peptide. Model of the full length OGT-UDP-peptide structure assembled from our complex structure (PDB code 3PE4) and the OGT TPR structure (PDB code 1W3B). (TXT 1035 kb)

Supplementary Model 3

PDB coordinates for the model of UDP-GlcNAc docked into hOGT4.5. UDP-GlcNAc was docked into the OGT-UDP structure (see Supplementary Fig. 5). (TXT 884 kb)

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Lazarus, M., Nam, Y., Jiang, J. et al. Structure of human O-GlcNAc transferase and its complex with a peptide substrate. Nature 469, 564–567 (2011). https://doi.org/10.1038/nature09638

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