Characterization of galactosyltransferase and sialyltransferase genes mediating the elongation of the extracellular O-GlcNAc glycans

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O-GlcNAc is a unique post-translational modification found in cytoplasmic, nuclear, and mitochondrial proteins. In a limited number of extracellular proteins, O-GlcNAc modifications occur through the action of EOGT, which specifically modifies subsets of epidermal growth factor-like (EGF) domain-containing proteins such as Notch receptors. The abnormalities due to EOGT mutations in mice and humans and the increased EOGT expression in several cancers signify the importance of EOGT pathophysiology and extracellular O-GlcNAc. Unlike intracellular O-GlcNAc monosaccharides, extracellular O-GlcNAc extends to form elongated glycan structures. However, the enzymes involved in the O-GlcNAc glycan extension have not yet been reported. In our study, we comprehensively screened potential galactosyltransferase and sialyltransferase genes related to the canonical O-GlcNAc glycan pathway and revealed the essential roles of B4GALT1 and ST3GAL4 in O-GlcNAc glycan elongation in human HEK293 cells. These findings were confirmed by sequential glycosylation of Drosophila EGF20 in vitro by EOGT, β4GalT-1, and ST3Gal-IV. Thus, the findings from our study throw light on the specific glycosyltransferases that mediate O-GlcNAc glycan elongation in human HEK293 cells.
OriginalsprogEngelsk
Artikelnummer149610
TidsskriftBiochemical and Biophysical Research Communications
Vol/bind703
Antal sider6
ISSN0006-291X
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
We thank K. Taki (Division for Medical Research Engineering, Nagoya University Graduate School of Medicine) for assistance with LC-MS/MS analysis, A. Ido (Gifu University) for assistance with in vitro glycosylation assay, M. Ogawa (Nagoya University) for supervision during the initial phase of the project, Y Kondo (Nagoya University) for critical comments and suggestions, and K. Moremen (University of Georgia) and the Repository of Glycoenzyme Expression Constructs (http://glycoenzymes. ccrc.uga.edu/) (the National Institutes of Health Grant P41GM103390 and P01GM107012) for the glycosyltransferase constructs. This work was supported by grants from the Japan Society for the Promotion of Science (JP23KJ1062 to YoT; JP19KK0195 and JP19H03176 to HT; JP19H03416 and 22H02815 to TO), Takeda Science Foundation (to HT), and Danish National Research Foundation (DNRF107 to CH), the Novo Nordisk Foundation (to CH).

Funding Information:
We thank K. Taki (Division for Medical Research Engineering, Nagoya University Graduate School of Medicine) for assistance with LC-MS/MS analysis, A. Ido (Gifu University) for assistance with in vitro glycosylation assay, M. Ogawa (Nagoya University) for supervision during the initial phase of the project, Y Kondo (Nagoya University) for critical comments and suggestions, and K. Moremen ( University of Georgia ) and the Repository of Glycoenzyme Expression Constructs ( http://glycoenzymes . ccrc.uga.edu/) (the National Institutes of Health Grant P41GM103390 and P01GM107012 ) for the glycosyltransferase constructs. This work was supported by grants from the Japan Society for the Promotion of Science ( JP23KJ1062 to YoT; JP19KK0195 and JP19H03176 to HT; JP19H03416 and 22H02815 to TO), Takeda Science Foundation (to HT), and Danish National Research Foundation ( DNRF107 to CH), the Novo Nordisk Foundation (to CH).

Publisher Copyright:
© 2024 Elsevier Inc.

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