56
IRUS Total
Downloads
  Altmetric

4-Deoxy-4-fluoro-GalNAz (4FGalNAz) is a metabolic chemical reporter of O-GlcNAc modifications, highlighting the notable substrate flexibility of O-GlcNAc transferase

Title: 4-Deoxy-4-fluoro-GalNAz (4FGalNAz) is a metabolic chemical reporter of O-GlcNAc modifications, highlighting the notable substrate flexibility of O-GlcNAc transferase
Authors: Jackson, EG
Cutolo, G
Yang, B
Yarravarapu, N
Burns, MWN
Bineva-Todd, G
Roustan, C
Thoden, JB
Lin-Jones, HM
Van Kuppevelt, TH
Holden, HM
Schumann, B
Kohler, JJ
Woo, CM
Pratt, MR
Item Type: Journal Article
Abstract: Bio-orthogonal chemistries have revolutionized many fields. For example, metabolic chemical reporters (MCRs) of glycosylation are analogues of monosaccharides that contain a bio-orthogonal functionality, such as azides or alkynes. MCRs are metabolically incorporated into glycoproteins by living systems, and bio-orthogonal reactions can be subsequently employed to install visualization and enrichment tags. Unfortunately, most MCRs are not selective for one class of glycosylation (e.g., N-linked vs O-linked), complicating the types of information that can be gleaned. We and others have successfully created MCRs that are selective for intracellular O-GlcNAc modification by altering the structure of the MCR and thus biasing it to certain metabolic pathways and/or O-GlcNAc transferase (OGT). Here, we attempt to do the same for the core GalNAc residue of mucin O-linked glycosylation. The most widely applied MCR for mucin O-linked glycosylation, GalNAz, can be enzymatically epimerized at the 4-hydroxyl to give GlcNAz. This results in a mixture of cell-surface and O-GlcNAc labeling. We reasoned that replacing the 4-hydroxyl of GalNAz with a fluorine would lock the stereochemistry of this position in place, causing the MCR to be more selective. After synthesis, we found that 4FGalNAz labels a variety of proteins in mammalian cells and does not perturb endogenous glycosylation pathways unlike 4FGalNAc. However, through subsequent proteomic and biochemical characterization, we found that 4FGalNAz does not widely label cell-surface glycoproteins but instead is primarily a substrate for OGT. Although these results are somewhat unexpected, they once again highlight the large substrate flexibility of OGT, with interesting and important implications for intracellular protein modification by a potential range of abiotic and native monosaccharides.
Issue Date: 21-Jan-2022
Date of Acceptance: 3-Dec-2021
URI: http://hdl.handle.net/10044/1/102003
DOI: 10.1021/acschembio.1c00818
ISSN: 1554-8929
Publisher: American Chemical Society
Start Page: 159
End Page: 170
Journal / Book Title: ACS Chemical Biology
Volume: 17
Issue: 1
Publication Status: Published
Online Publication Date: 2021-12-21
Appears in Collections:Chemistry



Unless otherwise indicated, items in Spiral are protected by copyright and are licensed under a Creative Commons Attribution NonCommercial NoDerivatives License.

Creative Commons