Tools for investigating O-GlcNAc in signaling and other fundamental biological pathways

GlycoID Proximity Labeling to Identify O‐GlcNAcylated Protein Interactomes in Live Cells

“…Dynamic O-GlcNAc glycosylation in mammalian cells plays a critical role in biological functions including stress responses (Martinez et al, 2017), nutrient sensing (Hart, 2019), and signaling (Nelson et al, 2024). Established tools to discover O-GlcNAc functions tend to perturb the cell environment, possibly through (1) metabolic engineering of the hexosamine pathway using reporter compounds like azide-labeled GlcNAz (Boyce et al, 2011) or 6-azido-GlcNAc (Chuh et al, 2014) analogs, among many other probes (Fehl & Hanover, 2022); (2) by labeling O-GlcNAc proteins in vitro using a chemoenzymatic strategy (Khidekel et al, 2003;Li et al, 2019), which maintains physiological O-GlcNAc patterns but loses spatial cellular context; or (3) histological O-GlcNAc staining using fixed and permeabilized cells (Aguilar et al, 2017), which loses the capacity for dynamic cellular responses.…”

“…Crucially, O-GlcNAc transiently regulates the function, localization, and stability of over 7000 intracellular human proteins (Wulff-Fuentes et al, 2021). Many signaling pathways are affected by O-GlcNAc, including metabolic homeostasis regulated by the hormones insulin and glucagon (Nelson et al, 2024). Insulin stimulation increases glucose uptake into cells while decreasing glucose synthesis, maintaining cellular glucose balance.…”