skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase

Authors:
; ; ; ORCiD logo; ORCiD logo; ; ; ; ; ORCiD logo; ORCiD logo;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1419061
Resource Type:
Journal Article
Resource Relation:
Journal Name: Nature Chemical Biology; Journal Volume: 13; Journal Issue: 12
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Hu, Chia-Wei, Worth, Matthew, Fan, Dacheng, Li, Baobin, Li, Hao, Lu, Lei, Zhong, Xiaofang, Lin, Ziqing, Wei, Liming, Ge, Ying, Li, Lingjun, and Jiang, Jiaoyang. Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase. United States: N. p., 2017. Web. doi:10.1038/nchembio.2494.
Hu, Chia-Wei, Worth, Matthew, Fan, Dacheng, Li, Baobin, Li, Hao, Lu, Lei, Zhong, Xiaofang, Lin, Ziqing, Wei, Liming, Ge, Ying, Li, Lingjun, & Jiang, Jiaoyang. Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase. United States. doi:10.1038/nchembio.2494.
Hu, Chia-Wei, Worth, Matthew, Fan, Dacheng, Li, Baobin, Li, Hao, Lu, Lei, Zhong, Xiaofang, Lin, Ziqing, Wei, Liming, Ge, Ying, Li, Lingjun, and Jiang, Jiaoyang. 2017. "Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase". United States. doi:10.1038/nchembio.2494.
@article{osti_1419061,
title = {Electrophilic probes for deciphering substrate recognition by O-GlcNAc transferase},
author = {Hu, Chia-Wei and Worth, Matthew and Fan, Dacheng and Li, Baobin and Li, Hao and Lu, Lei and Zhong, Xiaofang and Lin, Ziqing and Wei, Liming and Ge, Ying and Li, Lingjun and Jiang, Jiaoyang},
abstractNote = {},
doi = {10.1038/nchembio.2494},
journal = {Nature Chemical Biology},
number = 12,
volume = 13,
place = {United States},
year = 2017,
month =
}
  • The essential mammalian enzyme O-linked {beta}-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 sensor. OGT catalyses the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine (UDP-GlcNAc) to serines and threonines of cytoplasmic, nuclear and mitochondrial proteins, including numerous transcription factors, tumour suppressors, kinases, phosphatases and histone-modifying proteins. Aberrant glycosylation by OGT has been linked to insulin resistance, diabetic complications, cancer and neurodegenerative diseases including Alzheimer's. Despite the importance of OGT, the details of how it recognizes and glycosylates its protein substrates are largely unknown. We reportmore » here two crystal structures of human OGT, as a binary complex with UDP (2.8 {angstrom} resolution) and as a ternary complex with UDP and a peptide substrate (1.95 {angstrom}). 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.« less
  • O-Linked N-Acetylglucosamine (O-GlcNAc) is a reversible post-translational modification of Ser and Thr residues on cytosolic and nuclear proteins found in all higher eukaryotes. Aberrant O-GlcNAc modification of brain proteins has been linked to Alzheimer's disease (AD). However, understanding specific functions of O-GlcNAcylation in AD has been impeded by the difficulty in characterization of O-GlcNAc sites on proteins. In this study, we modified a chemical/enzymatic photochemical cleavage approach for enriching O-GlcNAcylated peptides in samples containing {approx}100 {micro}g of tryptic peptides from mouse cerebrocortical brain tissue. A total of 274 O-GlcNAcylated proteins were identified. Of these 168 were not previously known tomore » be modified by O-GlcNAc. Overall, 458 O-GlcNAc sites on Ser and Thr residues in 195 proteins were identified. Many of the modified residues are either known phosphorylation sites or located in close proximity to known phosphorylation sites. These findings support the proposed regulatory crosstalk between O-GlcNAcylation and phosphorylation. This study produced the most comprehensive O-GlcNAc proteome of mammalian brain tissue with both protein identification and O-GlcNAc site assignment. Interestingly, we observed O-{beta}-GlcNAc on EGF-like repeats in the extracellular domains of five membrane proteins, thus representing the first evidence for extracellular O-GlcNAcylation in mammalian systems by the ER-resident O-GlcNAc transferase (EOGT). We also report a GlcNAc-{beta}-1,3-Fuc-{alpha}-1-O-Thr modification on the EGF-like repeat of the Versican core protein, a novel substrate of Fringe {beta}1,3-N-acetylglucosaminyltransferases.« less