Use of metabolic glycoengineering and pharmacological inhibitors to assess lipid and protein sialylation on cells

Kranaster, Ramon; Blum, John D.; Dold, Jeremias E. G. A.; Wittmann, Valentin; Leist, Marcel

doi:10.1111/jnc.15737

Cited by 1 publication

(1 citation statement)

References 71 publications

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“…In the early years of MGE, most experiments were performed with human cancer or rodent cell lines, which readily accommodate sugar analog incorporation (Du et al., 2009; Keppler et al., 2001). Although a simplistic explanation, we observed that facile MGE labeling of cells was correlated with rapid cell proliferation (Jones et al., 2004; Yarema, 2002), which requires constant production of membrane components that host metabolically labeled glycans in both glycoproteins and glycolipids (Kayser, Geilen et al., 1992; Kranaster et al., 2022). Over time, MGE has expanded on many fronts to augment cell behavior, target intracellular and even extracellular components such as extracellular vesicles, serve as imaging agents, and improve the production of therapeutic proteins (Agatemor et al., 2019; Buettner et al., 2018; Dammen‐Brower et al., 2022).…”

Section: Strategic Planningmentioning

confidence: 99%

Protocol Considerations for In Vitro Metabolic Glycoengineering of Non‐Natural Glycans

et al. 2023

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Metabolic glycoengineering (MGE) refers to a technique where non-natural monosaccharide analogs are introduced into living biological systems. Once inside a cell, these compounds intercept a targeted biosynthetic glycosylation pathway and in turn are metabolically incorporated into cell-surface-displayed oligosaccharides, where they can modulate a host of biological activities or be exploited as tags for bioorthogonal and chemoselective ligation reactions. Over the past decade, azido-modified monosaccharides have become the go-to analogs for MGE; at the same time, analogs with novel chemical functionalities continue to be developed. Therefore, one emphasis of this article is to describe a general approach for analog selection and then provide protocols to ensure safe and efficacious analog usage by cells. Once cell-surface glycans have been successfully remodeled by MGE methodology, the stage is set for probing changes to the myriad cellular responses modulated by these versatile molecules. This manuscript concludes by detailing how one of these detection methods-flow cytometry-can be successfully utilized to quantify MGE analog incorporation and set the stage for numerous follow-up applications.

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Section: Strategic Planningmentioning

confidence: 99%

Protocol Considerations for In Vitro Metabolic Glycoengineering of Non‐Natural Glycans

et al. 2023

View full text Add to dashboard Cite

show abstract

Use of metabolic glycoengineering and pharmacological inhibitors to assess lipid and protein sialylation on cells

Abstract: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

Cited by 1 publication

References 71 publications

Protocol Considerations for In Vitro Metabolic Glycoengineering of Non‐Natural Glycans

Protocol Considerations for In Vitro Metabolic Glycoengineering of Non‐Natural Glycans

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