Click chemistry in sphingolipid research

Izquierdo, E.; Delgado, Antonio

doi:10.1016/j.chemphyslip.2018.07.004

Cited by 18 publications

(15 citation statements)

References 112 publications

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“…The most direct biochemical approaches have utilized mostly short chain ceramide species bound to agarose beads for either affinity chromatography columns or in tandem pull‐down/mass spectrometry studies 16 . Other assays use functionalized ceramides that may be cross‐linked with docking proteins to functionalized beads 17,18 . Together, these classic studies have found many associated proteins that bind ceramide and may be activated or inhibited, but still lacking the context on cellular topology.…”

Section: Introductionmentioning

confidence: 99%

Ceramide launches an acute anti‐adhesion pro‐migration cell signaling program in response to chemotherapy

et al. 2020

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Chemotherapy has been reported to upregulate sphingomylinases and increase cellular ceramide, often linked to the induction to cell death. In this work, we show that sublethal doses of doxorubicin and vorinostat still increased cellular ceramide, which was located predominantly at the plasma membrane. To interrogate possible functions of this specific pool of ceramide, we used recombinant enzymes to mimic physiological levels of ceramide at the plasma membrane upon chemotherapy treatment. Using mass spectrometry and network analysis, followed by experimental confirmation, the results revealed that this pool of ceramide acutely regulates cell adhesion and cell migration pathways with weak connections to commonly established ceramide functions (eg, cell death). Neutral sphingomyelinase 2 (nSMase2) was identified as responsible for the generation of plasma membrane ceramide upon chemotherapy treatment, and both ceramide at the plasma membrane and nSMase2 were necessary and sufficient to mediate these “side” effects of chemotherapy on cell adhesion and migration. This is the first time a specific pool of ceramide is interrogated for acute signaling functions, and the results define plasma membrane ceramide as an acute signaling effector necessary and sufficient for regulation of cell adhesion and cell migration under chemotherapeutical stress.

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Section: Introductionmentioning

confidence: 99%

Ceramide launches an acute anti‐adhesion pro‐migration cell signaling program in response to chemotherapy

et al. 2020

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“…To efficiently track the fate of dietary sphingolipids and identify sphingolipid-interacting microbial signatures, we leveraged a burgeoning tool -bioorthogonal click-chemistry -to investigate the trafficking and metabolism of sphingolipids. This approach offers high chemoselectivity and fast processing of broad sample types (40)(41)(42), making it an appealing tool to answer questions in host-microbe interactions. For example, copper (I) catalyzed azides alkynes cycloadditions are regarded as the archetype of click-chemistry (43), and are especially useful for our application.…”

Section: Introductionmentioning

confidence: 99%

Dietary sphinganine is selectively assimilated by members of the gut microbiome

Lee

Johnson

2020

Preprint

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Functions of the gut microbiome have a growing number of implications for host metabolic health, with diet being one of the most significant influences on microbiome composition. Compelling links between diet and the gut microbiome suggest key roles for various macronutrients, including lipids, yet how individual classes of dietary lipids interact with the microbiome remain largely unknown. A class of lipids known as sphingolipids are bioactive components of most foods and are produced by prominent gut microbes. This makes sphingolipids intriguing candidates for shaping diet-microbiome interactions.Here, we use a click-chemistry based approach to track the incorporation of bioorthogonal dietary omegaalkynyl sphinganine (sphinganine alkyne -SAA) into the gut microbial community (Click). Identification of microbe and SAA-specific metabolic products was achieved by fluorescence-based sorting of SAA containing microbes (Sort), 16S rRNA gene sequencing to identify the sphingolipid-interacting microbes (Seq), and comparative metabolomics to identify products of SAA assimilation by the microbiome (Spec). Together this approach, Click-Sort-Seq-Spec (ClickSSS), revealed that SAA-assimilation was nearly exclusively performed by gut Bacteroides, indicating that sphingolipid-producing bacteria play a major role in processing dietary sphinganine. Comparative metabolomics of cecal microbiota from SAAtreated mice showed conversion of SAA to a suite of dihydroceramides, consistent with metabolic activity via Bacteroides and Bifidobacterium. Additionally, other sphingolipid-interacting microbes were identified with a focus on an uncharacterized ability of Bacteroides and Bifidobacterium to metabolize 3 dietary sphingolipids. Therefore, ClickSSS provides a platform to study the flux of virtually any alkynelabeled metabolite in diet-microbiome interactions. 7 specific qualities of the alkyne, SAA-derived metabolites are revealed through the use of liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) (Spec).Application of Click-Sort-Seq-Spec (ClickSSS) (Fig. 1) revealed a set of microbes that take up dietary sphingolipids. This included the sphingolipid-producing Bacteroides and beneficial microbes such as Bifidobacterium that do not produce sphingolipids. Comparative metabolomics revealed the dietdependent sphingolipidome of both Bacteroides thetaiotaomicaron (B. theta) and Bifidobacterium longum subsp. infantis (B. longum). Our analysis suggests that diets rich in sphingolipids can influence the composition of the microbiome with implications for supporting the colonization of beneficial microbes. Moreover, ClickSSS can be applied to a variety of diet-microbiome systems to uncover the mechanism of nutrient interactions with the gut microbial metabolome.8 Materials and Methods General InformationAll buffer, reagents, media, and instrumentation were utilized sterile. All solvents used for metabolomics were purchased from Fisher Scientific as HPLC grade. Bacterial culturingBacteroides thetaiotaomicron strain V...

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“…[2] In addition, the 1,3-dipolar nature of azides has been exploited in click chemistry as a convenient method for the synthesis of glycoconjugates from glycosyl azides [3] and to append biomarkers to macromolecules. [4] Specifically, the azidation of benzhydryl alcohols is targeted for the preparation of azidodiarylmethanes as precursors to (diarylmethyl)amines, which are prominent constituents of commercial pharmaceuticals, notably, cetirizine and letrozole ( Figure 1). Aside from their synthetic versatility, azides have also demonstrated activity as pharmacophores, such as in the HIV/ AIDS drug zidovudine.…”

Section: Introductionmentioning

confidence: 99%

“…Azides are recognized in synthesis as valuable precursors to amines, nitrenes and heterocycles . In addition, the 1,3‐dipolar nature of azides has been exploited in click chemistry as a convenient method for the synthesis of glycoconjugates from glycosyl azides and to append biomarkers to macromolecules . Specifically, the azidation of benzhydryl alcohols is targeted for the preparation of azidodiarylmethanes as precursors to (diarylmethyl)amines, which are prominent constituents of commercial pharmaceuticals, notably, cetirizine and letrozole (Figure ).…”

Section: Introductionmentioning

confidence: 99%

A Direct Brønsted Acid‐Catalyzed Azidation of Benzhydrols and Carbohydrates

2019

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Benzhydryl alcohols were converted into their corresponding diarylazidomethane analogues using azidotrimethylsilane (TMSN3) in the presence of a catalytic amount of a Brønsted acid HBF4·OEt2. The azidation reactions proceeded in high yields and demonstrated excellent functional group tolerance to electron‐donating and electron‐withdrawing substituents. In addition, a range of unprotected functional groups including amine, amide, aldehyde and alcohol were well‐tolerated. Furthermore, this methodology was successfully applied to carbohydrates for the preparation of the corresponding azide derivatives.

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Click chemistry in sphingolipid research

Cited by 18 publications

References 112 publications

Ceramide launches an acute anti‐adhesion pro‐migration cell signaling program in response to chemotherapy

Ceramide launches an acute anti‐adhesion pro‐migration cell signaling program in response to chemotherapy

Dietary sphinganine is selectively assimilated by members of the gut microbiome

A Direct Brønsted Acid‐Catalyzed Azidation of Benzhydrols and Carbohydrates

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