Introduction to Glycosylation Inhibition Assays
Glycosylation is a crucial post-translational modification that influences protein folding, stability, trafficking, and cellular signaling. Disrupting glycosylation pathways has become a valuable strategy for probing glycan-dependent processes and developing targeted therapeutics. Creative Biolabs offers glycosylation inhibition assays that use chemical or genetic tools to block the function of glycosyltransferases (GTs) or associated enzymes responsible for transferring sugar moieties from activated donors to proteins or lipids.
Key Inhibitory Strategies
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Strategy
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Description
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Substrate Analogues
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Structural mimics of natural sugar donors, such as 2-fluorofucose, to disrupt fucosylation.
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Metabolic Blockade
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Compounds like tunicamycin block nucleotide sugar synthesis.
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Catalytic Inhibition
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Direct inhibitors that bind to GT active sites, such as kifunensine inhibiting mannosidase I.
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Our Glycosylation Inhibitor Portfolio
Creative Biolabs offers a comprehensive selection of glycosylation inhibitors that target different biosynthetic routes. These inhibitors support studies in cancer biology, infectious disease, and immunology.
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Inhibitor Class
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Representative Compounds
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Target Pathway
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Applications
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N-Glycosylation
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Tunicamycin, Kifunensine, Swainsonine
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OST complex, ER mannosidases
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Protein folding, tumor glycan profiling
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O-Glycosylation
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Ac5GalNTGc, T3Inh-1, Luteolin
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GALNTs, Core-1 synthase
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Cancer migration, Tn antigen expression
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GAG Biosynthesis
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4-Methylumbelliferone (4-MU)
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Hyaluronan synthase
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Inflammatory regulation, tissue repair
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GSL Synthesis
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NB-DNJ, PDMP
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Glucosylceramide synthase
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Lysosomal storage disorder studies
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GPI Anchor Formation
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Mannosamine analogues
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Phosphoethanolamine transferases
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GPI-anchored protein shedding
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O-GlcNAcylation
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OSMI-1, Thiamet-G
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OGT and OGA
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Signal transduction modulation
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Case Spotlight: Tunicamycin and N-Glycosylation Inhibition
Tunicamycin blocks GlcNAc-1-phosphate transferase (GPT), halting the synthesis of dolichol-linked oligosaccharides and preventing N-glycosylation at the ER. This induces ER stress and disrupts proper protein folding, making it a key tool in:
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Investigating folding quality control
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Enhancing cancer cell apoptosis
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Studying immune checkpoint glycosylation
Enzyme Engineering for Glycosylation Modulation
Our enzyme engineering services allow precise disruption of glycosylation via rationally designed glycosyltransferase variants:
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Catalytically inactive mutants
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Substrate-swapped GTs with altered specificity
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Stable knock-in systems using gene-editing techniques or plasmid expression
Advanced Analytical Technologies
To accurately assess glycosylation inhibition outcomes, we integrate multiple high-resolution readouts:
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Technique
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Description
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Outcome
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High-Throughput Screening (HTS)
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Fluorescent/luminescent assays for rapid inhibitor screening
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Inhibitor potency ranking
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Western Blot & Click Chemistry
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Detection with anti-glycan antibodies or azido-labeled probes
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Glycan-specific visualization
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Mass Spectrometry (LC-MS/MS)
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Site-specific glycoproteomics and glycomics
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Structural profiling of glycan changes
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Flow Cytometry
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Quantification of surface glycans
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Cell population analysis
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Why Choose Creative Biolabs
We offer an unmatched combination of technical depth, flexible service options, and expert consulting:
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Broad spectrum of glycosylation inhibitors
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Integrated analytical platform with mass spectrometry, lectin profiling, and flow cytometry
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Tailored workflows for compound screening, site-mapping, and cell-based assays
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Optional enzyme engineering and custom GT mutant development
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End-to-end project support, from study design to data analysis
Ready to explore glycosylation inhibition for your project? Whether you're screening inhibitors, validating glycoengineered therapies, or dissecting glycan function in disease models, our experts are here to help. Contact us for your tailor solutions of glycosylation inhibition research.
FAQs
Q: What types of glycosylation can your inhibition services target?
A: At Creative Biolabs, we offer targeted inhibition services for both N-linked and O-linked glycosylation, as well as C-glycosylation and GPI-anchor biosynthesis upon request. Our platform employs a suite of small molecule inhibitors, and gene knockout to selectively inhibit key enzymes such as oligosaccharyltransferases, mannosidases, fucosyltransferases, sialyltransferases, and UDP-sugar transporters. This allows us to study glycan-dependent processes like protein folding, immune recognition, metastasis, and receptor activation with pathway-level precision.
Q: How do you ensure specificity and minimize off-target effects in glycosylation inhibition?
A: Specificity is addressed at multiple levels. First, we use well-validated inhibitors with known IC50 and Ki values against their target glycosyltransferases or glycosidases. Second, we apply dose-response titration and time-course analysis to optimize concentration and exposure windows. Third, we use orthogonal readouts, including glycoprotein profiling (e.g., via LC-MS/MS, lectin blotting), transcriptomics, and viability assays to confirm functional inhibition with minimal cytotoxicity. When available, we also provide rescue experiments with enzyme overexpression or sugar supplementation to verify pathway-specific effects.
Q: What types of samples or cell lines can be used in your glycosylation inhibition studies?
A: Our services are compatible with a wide range of mammalian cell lines. We also support glycoengineering projects in bacterial, yeast, or insect systems, provided that the glycosylation pathways are characterized. Clients can supply their own cell lines or work with our pre-established validated models. For in vivo studies, we offer protocol development support and can provide inhibitor formulations suitable for murine models.
Reference
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Almahayni, Karim, et al. "Small molecule inhibitors of mammalian glycosylation." Matrix Biology Plus 16 (2022): 100108. https://doi.org/10.1016/j.mbplus.2022.100108
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