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PreprintCited by 17Glycosylation and Glycoproteins ResearchCarbohydrate Chemistry and SynthesisProtein Structure and Dynamics

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Exploration of O-GlcNAc-transferase (OGT) glycosylation sites reveals a target sequence compositional bias

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Chong, P. Andrew, Nosella, Michael, Vanama, Manasvi, Ruiz-Arduengo, Roxana, Forman-Kay, Julie D.·2022·Journal of Biological Chemistry
DOI 10.1101/2022.09.12.507593PMC PMC10164912

Preprint — published as 10.1016/j.jbc.2023.104629

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Abstract

O-GlcNAc transferase (OGT) is an essential glycosylating enzyme that catalyzes the addition of N-acetylglucosamine to serine or threonine residues of nuclear and cytoplasmic proteins. The enzyme glycosylates a broad range of peptide sequences and prediction of glycosylation sites has proven challenging. The lack of an experimentally verified set of polypeptide sequences that are not glycosylated by OGT has made prediction of legitimate glycosylation sites more difficult. Here, we tested a number of intrinsically disordered protein regions as substrates of OGT to establish a set of sequences that are not glycosylated by OGT. The negative data set suggests an amino acid compositional bias for OGT targets. This compositional bias was validated by modifying the amino acid composition of the protein Fused in sarcoma (FUS) to enhance glycosylation. NMR experiments demonstrate that the tetratricopeptide repeat (TPR) region of OGT can bind FUS and that glycosylation-promoting mutations enhance binding. These results provide evidence that the TPR recognizes disordered segments of substrates with particular compositions to promote glycosylation, providing insight into the broad specificity of OGT.

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Metadata

Authors
Chong, P. A., Nosella, M., Vanama, M., Ruiz-Arduengo, R., & Forman-Kay, J. D.
Affiliations
  • Hospital for Sick Children (CA)
  • University of Toronto (CA)
Corresponding author
Forman-Kay, Julie D.
ORCID
Venue
Journal of Biological Chemistry
Publisher
openRxiv
Published
2022-09-13
Volume
299
Issue
5
Pages
104629-104629
ISSN
0021-9258 (ISSN-L)
DOI
10.1101/2022.09.12.507593
PMC
PMC10164912
OpenAlex
W4360942084
Cited by
17
References
1
Citations by year
2026: 6 · 2025: 6 · 2024: 3 · 2023: 2
FWCI
1.51
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Canadian Institutes of Health Research (148375, 148375_1)
  • Natural Sciences and Engineering Research Council of Canada (-2016-06718, 06718, -148375)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
O-GlcNAc transferase, O-GlcNAcylation, glycosylation site prediction, intrinsically disordered proteins, protein-protein binding, tetratricopeptide repeats, FUS, N-acetylglucosamine
MeSH terms
Golgi Matrix Proteins, Amino Acids, Glycosylation, Humans, Magnetic Resonance Imaging, Mutation, N-Acetylglucosaminyltransferases, Computational Biology, RNA-Binding Protein EWS, TATA-Binding Protein Associated Factors, Adaptor Proteins, Signal Transducing

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