Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells

Walter, Tim; Collenburg, Lena; Japtok, Lukasz; Kleuser, Burkhard; Schneider‐Schaulies, Sibylle; Müller, Nora; Bécam, Jérôme; Schubert‐Unkmeir, Alexandra; Kong, Ji Na; Bieberich, Erhard; Seibel, Jürgen

doi:10.1039/c6cc02879a

Cited by 23 publications

(21 citation statements)

References 19 publications

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“…Bifunctional or biorthogonal lipid analogues contain a diazirine group for UV crosslinking of the analogue to a binding protein and an alkyne group for derivatization with fluorophores or biotin using click chemistry (Haberkant and van Meer, 2009;Haberkant et al, 2013;Haberkant and Holthuis, 2014). In another approach, azido-derivatized ceramide and ceramide analogues (S18) have been used with click chemistry to label ceramide-binding proteins (e.g., azido-S18 interaction with aPKC) in living cells (Walter et al, 2016). In addition, bifunctional lipid analogues have been used to identify or visualize fatty acid, cholesterol, sphingosine, and ceramide binding proteins and will certainly allow for a similar task with lysosphingolipids such as psychosine (Haberkant et al, 2013(Haberkant et al, , 2016Hulce et al, 2013;Kong et al, 2015).…”

Section: Next-generation Lipid Analysismentioning

confidence: 99%

Lysosphingolipids and sphingolipidoses: Psychosine in Krabbe's disease

Spassieva

Bieberich

2016

J of Neuroscience Research

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Until recently, lipids were considered inert building blocks of the cellular membranes. This changed three decades ago when lipids were found to regulate cell polarity and vesicle transport, and the “lipid raft” concept took shape. The lipid-driven membrane anisotropy in form of “rafts” that associate with proteins led to the view that organized complexes of lipids and proteins regulate various cell functions. Disturbance of this organization can lead to cellular, tissue, and organ malfunction. Sphingolipidoses, lysosomal storage diseases that are caused by enzyme deficiencies in the sphingolipid degradation pathway, were found to be particularly detrimental to the brain. These enzyme deficiencies result in accumulation of sphingolipid metabolites in lysosomes, although it is not yet clear how this accumulation affects the organization of lipids in cellular membranes. Krabbe disease or globoid cell leukodystrophy was one of the first sphingolipidosis for which the raft concept offered a potential mechanism. Krabbe disease is caused by mutations in the enzyme β-galactocerebrosidase, however, elevation of its substrate, galactosylceramide, is not observed or considered detrimental. Instead, it was found that a byproduct of galactosylceramide metabolism, the lysosphingolipid psychosine, is accumulated. The “psychosine hypothesis” has been refined by showing that psychosine disrupts “lipid rafts” and vesicular transport critical for the function of glia and neurons. The role of psychosine in Krabbe disease is an example of how the disruption of the sphingolipid metabolism can lead to elevation of a toxic lysosphingolipids resulting in disruption of cellular membrane organization and neurotoxicity.

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Section: Next-generation Lipid Analysismentioning

confidence: 99%

Lysosphingolipids and sphingolipidoses: Psychosine in Krabbe's disease

Spassieva

Bieberich

2016

J of Neuroscience Research

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“…Ceramide-rich membrane areas promote structural changes within the plasma membrane, which segregate membrane receptors and affect the membrane curvature and vesicle formation, fusion and trafficking. 19,20 We selected ω-N3-C6-ceramide, which is efficiently incorporated into cellular membranes and can be click-labeled with DBCO-functionalized dyes for fluorescence imaging 21,22 , for further functionalization with a primary amino group (Supporting Information and Supplementary Figures S1-S13). Therefore, we synthesized α-NH2-ω-N3-C6-ceramide from (tert-butoxycarbonyl)-Llysine ( Figure 1A).…”

Section: Resultsmentioning

confidence: 99%

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

Goetz

Kunz

Fink

et al. 2020

Preprint

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Expansion microscopy (ExM) enables super-resolution imaging of proteins and nucleic acids on conventional microscopes. However, imaging of details of the organization of lipid bilayers by light microscopy remains challenging. We introduce an azide-and amino-modified sphingolipid ceramide, which upon incorporation into membranes can be labeled by click chemistry and linked into hydrogels, followed by 4x to 10x expansion. Confocal and structured illumination microscopy (SIM) enabled imaging of sphingolipids and their interactions with proteins in the membrane of intracellular organelles with a spatial resolution of 10-20 nm. Because sphingolipids accumulated efficiently in pathogens we used sphingolipid ExM to investigate bacterial infections of human HeLa229 cells by Neisseria gonorrhoeae, Chlamydia trachomatis and Simkania negevensis with a resolution so far only provided by electron microscopy. In particular, sphingolipid ExM allowed us to visualize the inner and outer membrane of intracellular bacteria and determine their distance to 27.6 ± 7.7 nm. The Supporting Information is available free of charge on the ACS Publications website. Supplementary Figures S1-S23. Supplementary Movie 1: 10x ExM SIM z-stack of Hela229 cells infected with Chlamydia trachomatis for 24 h, fed with α-NH2-ω-N3-C6-ceramide, fixed, permeabilized and stained with DBCO-Alexa Fluor 488. Chlamydia are clearly located at the inclusion membrane. Scale bar, 10 µm.

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“…For example, feeding the cells with severaln ew ceramide analoguesh ave provedt o be an ovel alternative for cell surface modifications, which enables the visualizationo ft he plasma membrane of Jurkat and human brain microvascular endothelial cells by conjugation with fluorescent compounds via click chemistry. [91,92] In the past decades, the direct chemicalm odification approach offers an easy way to functionalize the cell surface withouta ny genetic engineering of the cells, but suffersf rom the problem of off-site modifications. The recently developed affinity labelling chemistry holds great potentialf or better controlled reactions.…”

Section: Discussionmentioning

confidence: 99%

Chemical and Enzymatic Strategies for Bacterial and Mammalian Cell Surface Engineering

Yin

Guanbang

et al. 2018

Chemistry A European J

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The cell surface serves important functions such as the regulation of cell-cell and cell-environment interactions. The understanding and manipulation of the cell surface is important for a wide range of fundamental studies of cellular behavior and for biotechnological and medical applications. With the rapid advance of biology, chemistry and materials science, many strategies have been developed for the functionalization of bacterial and mammalian cell surfaces. Here, we review the recent development of chemical and enzymatic approaches to cell surface engineering with particular emphasis on discussing the advantages and limitations of each of these strategies.

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Incorporation and visualization of azido-functionalized N-oleoyl serinol in Jurkat cells, mouse brain astrocytes, 3T3 fibroblasts and human brain microvascular endothelial cells

Abstract: The synthesis and biological evaluation of azido-N-oleoyl serinol is reported. It mimicks biofunctional lipid ceramides and has shown to be capable of click reactions for cell membrane imaging in Jurkat and human brain microvascular endothelial cells.

Cited by 23 publications

References 19 publications

Lysosphingolipids and sphingolipidoses: Psychosine in Krabbe's disease

Lysosphingolipids and sphingolipidoses: Psychosine in Krabbe's disease

Nanoscale imaging of bacterial infections by sphingolipid expansion microscopy

Chemical and Enzymatic Strategies for Bacterial and Mammalian Cell Surface Engineering

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