Synthesis of biolabile thioalkyl-protected phosphates from an easily accessible phosphotriester precursor

Willems, Lianne I.; Murphy, Lloyd Declan; Huxley, Kathryn; Wilding, Ava; Robinson, Cyane; Foucart, Quentin

doi:10.1039/d3sc00693j

Cited by 2 publications

(1 citation statement)

References 42 publications

(69 reference statements)

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“…48 While we and others have made sugar-1-phosphates caged as labile bis- S -acetylthioethyl (SATE) phosphotriesters, synthesis of SATE-caged 6AzGlc-1-phosphate failed in our hands. 42,49,50 Instead, we took inspiration from the increasingly popular protide technology that has gained attention to cage phosphates in antiviral nucleotides. 51,52 To our knowledge, such chemistry has not been applied to sugar-1-phosphates yet, but for instance to sugar-6-phosphates.…”

Section: Resultsmentioning

confidence: 99%

Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells

Li,

Di Vagno,

Chawla

et al. 2023

Preprint

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Mammalian cells orchestrate signalling through interaction events on their surfaces. Proteoglycans are an intricate part of these interactions, carrying large glycosaminoglycan polysaccharides that recruit signalling molecules. Despite their importance in development, cancer and neurobiology, a relatively small number of proteoglycans have been identified. In addition to the complexity of glycan extension, biosynthetic redundancy in the first protein glycosylation step by two xylosyltransferase isoenzymes XT1 and XT2 complicates annotation of proteoglycans. Here, we develop a chemical strategy that allows profiling of cellular proteoglycans. By employing a tactic termed bump-and-hole engineering, we engineer the two isoenzymes XT1 and XT2 to specifically transfer a chemically modified xylose analogue to target proteins. The chemical modification contains a bioorthogonal tag, allowing the ability to visualise and profile target proteins modified by both transferases in mammalian cells as well as pinpointing glycosylation sites by mass spectrometry. Engineered XT enzymes permit a view into proteoglycan biology that is orthogonal to conventional techniques in biochemistry.

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Section: Resultsmentioning

confidence: 99%

Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells

Li,

Di Vagno,

Chawla

et al. 2023

Preprint

Self Cite

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Probing Glycerolipid Metabolism using a Caged Clickable Glycerol‐3‐Phosphate Probe

Lou,

Ancajas,

Zhou

et al. 2024

ChemBioChem

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In this study, we present the probe SATE‐G3P‐N3 as a novel tool for metabolic labeling of glycerolipids (GLs) to investigate lipid metabolism in yeast cells. By introducing a clickable azide handle onto the glycerol backbone, this probe enables general labeling of glycerolipids. Additionally, this probe contains a caged phosphate moiety at the glycerol sn‐3 position to not only facilitate probe uptake by masking negative charge but also to bypass the phosphorylation step crucial for initiating phospholipid synthesis, thereby enhancing phospholipid labeling. The metabolic labeling activity of the probe was thoroughly assessed through cellular fluorescence microscopy, mass spectrometry (MS), and thin‐layer chromatography (TLC) experiments. Fluorescence microscopy analysis demonstrated successful incorporation of the probe into yeast cells, with labeling predominantly localized at the plasma membrane. LCMS analysis confirmed metabolic labeling of various phospholipid species (PC, PS, PA, PI, and PG) and neutral lipids (MAG, DAG, and TAG), and GL labeling was corroborated by TLC. These results showcased the potential of the SATE‐G3P‐N3 probe in studying GL metabolism, offering a versatile and valuable approach to explore the intricate dynamics of lipids in yeast cells.

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Synthesis of biolabile thioalkyl-protected phosphates from an easily accessible phosphotriester precursor

Abstract: Robust methods for the synthesis of mixed phosphotriesters are essential to accelerate the development of novel phosphate-containing bioactive molecules. To enable efficient cellular uptake, phosphate groups are commonly masked with...

Cited by 2 publications

References 42 publications

Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells

Xylosyltransferase Bump-and-hole Engineering to Chemically Manipulate Proteoglycans in Mammalian Cells

Probing Glycerolipid Metabolism using a Caged Clickable Glycerol‐3‐Phosphate Probe

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