A Coumarin Triflate Reagent Enables One‐Step Synthesis of Photo‐Caged Lipid Metabolites for Studying Cell Signaling

Wagner, Nicolai; Schuhmacher, Milena; Lohmann, Annett; Nadler, André

doi:10.1002/chem.201903909

Cited by 7 publications

(8 citation statements)

References 35 publications

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“…Photocaged lipids have emerged as attractive strategy for the photoactivation of lipids through cleavage of a photosensitive protection group to reveal the native lipid . Photocaged versions of phosphatidic acid , have been used to control the activation of matrix metalloproteinase 2 and flagellar excision . Chemical modifications of the photosensitive groups also allow for subcellular targeting. − …”

Section: Introductionmentioning

confidence: 99%

Optical Control of Phosphatidic Acid Signaling

et al. 2021

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Phosphatidic acids (PAs) are glycerophospholipids that regulate key cell signaling pathways governing cell growth and proliferation, including the mTOR and Hippo pathways. Their acyl chains vary in tail length and degree of saturation, leading to marked differences in the signaling functions of different PA species. For example, in mTOR signaling, saturated forms of PA are inhibitory, whereas unsaturated forms are activating. To enable rapid control over PA signaling, we describe here the development of photoswitchable analogues of PA, termed AzoPA and dAzoPA , that contain azobenzene groups in one or both lipid tails, respectively. These photolipids enable optical control of their tail structure and can be reversibly switched between a straight trans form and a relatively bent cis form. We found that cis - dAzoPA selectively activates mTOR signaling, mimicking the bioactivity of unsaturated forms of PA. Further, in the context of Hippo signaling, whose growth-suppressing activity is blocked by PA, we found that the cis forms of both AzoPA and dAzoPA selectively inhibit this pathway. Collectively, these photoswitchable PA analogues enable optical control of mTOR and Hippo signaling, and we envision future applications of these probes to dissect the pleiotropic effects of physiological and pathological PA signaling.

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Section: Introductionmentioning

confidence: 99%

Optical Control of Phosphatidic Acid Signaling

et al. 2021

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“…Recently, Schultz and co-workers reported a photocaged version of LPA that allowed for light-induced activation of LPA receptor-dependent effects, including chemotaxis . However, photoactivation of this probe is not reversible, and it has a chemically modified headgroup, which perturbs the amphiphilic character of the molecule and might affect trafficking. , In recent years, we and others have developed a series of photoswitchable lipids and demonstrated their capacity for the reversible optical control of lipid metabolism and signaling. These photoswitchable lipids have an azobenzene photoswitch incorporated into the hydrophobic tail and mediate optical control of lipid function by reversible, light-induced isomerization between the trans - (straight) and cis - (bent) isomers.…”

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confidence: 99%

Optical Control of Lysophosphatidic Acid Signaling

et al. 2020

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Lysophosphatidic acid (LPA) is a phospholipid that acts as an extracellular signaling molecule and activates the family of lysophosphatidic acid receptors (LPA 1−6 ). These G protein-coupled receptors (GPCRs) are broadly expressed and are particularly important in development as well as in the nervous, cardiovascular, reproductive, gastrointestinal, and pulmonary systems. Here, we report on a photoswitchable analogue of LPA, termed AzoLPA, which contains an azobenzene photoswitch embedded in the acyl chain. AzoLPA enables optical control of LPA receptor activation, shown through its ability to rapidly control LPA-evoked increases in intracellular Ca 2+ levels. AzoLPA shows greater activation of LPA receptors in its light-induced cis-form than its dark-adapted (or 460 nm light-induced) trans-form. AzoLPA enabled the optical control of neurite retraction through its activation of the LPA 2 receptor.

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“…Reagent stability was a concern since only a single functionalization reaction involving a benzylic triflate reagent had been reported before . Nevertheless, upon generation of DEAC-triflate in situ and direct use of the reaction mixture for the functionalization reaction, we found that native lipids could indeed be selectively equipped with a photocaging group at the phosphate or carboxylate moieties . We demonstrated the utility of this transformation for a number of potent G-protein-coupled receptor (GPCR) ligands, including lysophosphatidic acid, platelet activating factor, and prostaglandin E2.…”

Section: Main Textmentioning

confidence: 98%

“… 2019 25 15483 15487 . 3 Show the in situ synthesis of diethylaminocoumarin-triflate to directly functionalize native lipids with a photocaging group at the phosphate or carboxylate moieties. Live-Cell Lipid Biochemistry Reveals a Role of Diacylglycerol Side-Chain Composition for Cellular Lipid Dynamics and Protein Affinities Schuhmacher M. Grasskamp A. T. Barahtjan P. Wagner N. Lombardot B.…”

Section: Key Referencesmentioning

confidence: 99%

The Road to Quantitative Lipid Biochemistry in Living Cells

Iglesias-Artola

Nadler

2023

Acc. Chem. Res.

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Metrics & MoreArticle Recommendations CONSPECTUS: Traditional cell biological techniques are not readily suitable for studying lipid signaling events because genetic perturbations are much slower than the interconversion of lipids in complex metabolic networks. For this reason, novel chemical biological approaches have been developed. One approach is to chemically modify a lipid with a so-called "caging group" that renders it inactive, but this cage can be removed photochemically inside cells to release the bioactive molecule. These caged compounds offer unique advantages for studying the kinetics of cellular biochemistry and have been extensively used in the past. However, a limitation of conventional caged compounds is their ability to diffuse freely inside the cell, which does not permit localized activation below optical precision. This poses a challenge for studying lipid signaling as lipid function inside cells is tightly linked to their parent membrane. An ideal lipid probe should, therefore, be restricted to a single organelle membrane or preferentially to a single leaflet. We first demonstrated the plasma-membrane-specific photorelease of fatty acids by employing sulfonated caging groups. Using these caged fatty acid probes we demonstrated that lipid localization determines signaling outcome.Generalizing this approach, we designed a so-called "click cage" that can be coupled to lipids and offers the possibility to attach organelle targeting groups via click chemistry. Using this strategy, we have synthesized plasma membrane, lysosomal, mitochondria, and endoplasmic-reticulum-targeted lipids that can be used to dissect organelle-specific signaling events. To reduce the synthetic effort associated with generating caged compounds, we designed a coumarin triflate reagent that allows the direct functionalization of phosphate-or carboxylate-containing compounds. With this novel reagent, we synthesized a small library of photocaged G-proteincoupled receptor (GPCR) ligands to study the underlying lipid signaling dynamics. Most recently, we have focused on quantifying the kinetics of lipid signaling for different diacylglycerol (DAG) species using plasma-membrane-targeted caged DAGs. Using this approach, we quantitatively measured lipid−protein affinities and lipid transbilayer dynamics in living cells. After analyzing DAGs with different acyl chain length and saturation degree, we discovered that affinities can vary by up to an order of magnitude. This finding clearly shows that cells are able to distinguish between individual DAG species, thereby demonstrating that lipid diversity matters in cellular signal processing. Although the recent advances have yielded valuable tools to study lipid signaling, challenges remain on specifically targeting the different leaflets of organelle membranes. Furthermore, it is necessary to simplify the experimental approaches required for parametrizing and corroborating quantitative kinetic models of lipid signaling. In the future, we envision that the application of leaflet-spec...

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A Coumarin Triflate Reagent Enables One‐Step Synthesis of Photo‐Caged Lipid Metabolites for Studying Cell Signaling

Cited by 7 publications

References 35 publications

Optical Control of Phosphatidic Acid Signaling

Optical Control of Phosphatidic Acid Signaling

Optical Control of Lysophosphatidic Acid Signaling

The Road to Quantitative Lipid Biochemistry in Living Cells

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