Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates

Kuerschner, Lars; Leyendecker, Philipp; Klizaite, Kristina; Fiedler, Maria; Saam, Jennifer; Thiele, Christoph

doi:10.1016/j.jlr.2022.100188

Cited by 6 publications

(9 citation statements)

References 49 publications

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“…This end product has been found unstable ( Patel and Walt, 1988 ) and thus escaped accurate detection and quantification. To overcome this potential drawback, oxaalkyne FAs ( Figure 1B ) have recently been introduced as dedicated tracers to study β-oxidation ( Kuerschner et al, 2022 ). Here, an oxygen atom replaces a carbon atom within the hydrocarbon chain, and during catabolic shortening of the chain, the substituting oxygen eventually arrests further processing ( Figure 4B ).…”

Section: Alkyne Fatty Acids As Tracers For Lipid Catabolismmentioning

confidence: 99%

“…These metabolites may also fulfill a signaling function as they carry information about the metabolic status of this major β-oxidizing tissue, ref. ( Kuerschner et al, 2022 ).…”

Section: Alkyne Fatty Acids As Tracers For Lipid Catabolismmentioning

confidence: 99%

“…Using this procedure, a set of experiments was performed investigating lipid metabolism in hepatocytes in unprecedented detail ( Figure 3 , refs. Thiele et al, 2019 ; Wunderling et al, 2021 ; Kuerschner et al, 2022 ). Lipidomics data on the anabolism of palmitate and linoleate were collected using the alkyne tracers FA 17:0;Y or FA 18:2;Y.…”

Section: Introductionmentioning

confidence: 99%

“…(B,C) Compared to saturated long-chain FAs that were assimilated quite broadly into various lipid classes after 6 h (B) , medium-chain FAs labeled primarily the TG pool (C) , ref. ( Kuerschner et al, 2022 ). (D) Major part of medium-chain fatty acid TG (MCFA-TG) was synthesized by diacylglycerol acyltransferase 1 (DGAT1) whose activity can only be partially substituted by DGAT2, ref.…”

Section: Introductionmentioning

confidence: 99%

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Tracing Lipid Metabolism by Alkyne Lipids and Mass Spectrometry: The State of the Art

Kuerschner

Thiele

2022

Front. Mol. Biosci.

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Lipid tracing studies are a key method to gain a better understanding of the complex metabolic network lipids are involved in. In recent years, alkyne lipid tracers and mass spectrometry have been developed as powerful tools for such studies. This study aims to review the present standing of the underlying technique, highlight major findings the strategy allowed for, summarize its advantages, and discuss some limitations. In addition, an outlook on future developments is given.

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Section: Alkyne Fatty Acids As Tracers For Lipid Catabolismmentioning

confidence: 99%

“…These metabolites may also fulfill a signaling function as they carry information about the metabolic status of this major β-oxidizing tissue, ref. ( Kuerschner et al, 2022 ).…”

Section: Alkyne Fatty Acids As Tracers For Lipid Catabolismmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Tracing Lipid Metabolism by Alkyne Lipids and Mass Spectrometry: The State of the Art

Kuerschner

Thiele

2022

Front. Mol. Biosci.

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“…Compared to mass spectrometry-based techniques, fluorescence analysis using functionally optimized probes can facilitate sensitive and selective observation of metabolic activity without the need of special detection techniques and complicated data analysis. Despite these advantages, such fluorescence-based analyses are still scarce. − Although fluorescent probes for single metabolic reactions − and metabolites , have been reported to date, these probes do not allow flux analysis of an entire metabolic pathway.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence-Based Detection of Fatty Acid β-Oxidation in Cells and Tissues Using Quinone Methide-Releasing Probes

et al. 2023

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Detection of metabolic activity enables us to reveal the inherent metabolic state of cells and elucidate mechanisms underlying cellular homeostasis and growth. However, a fluorescence approach for the study of metabolic pathways is still largely unexplored. Herein, we have developed a new chemical probe for the fluorescence-based detection of fatty acid β-oxidation (FAO), a key process in lipid catabolism, in cells and tissues. This probe serves as a substrate of FAO and forms a reactive quinone methide (QM) as a result of metabolic reactions. The liberated QM is covalently captured by intracellular proteins, and subsequent bio-orthogonal ligation with a fluorophore enables fluorescence analysis. This reaction-based sensing allowed us to detect FAO activity in cells at a desired emission wavelength using diverse analytical techniques including fluorescence imaging, in-gel fluorescence activity-based protein profiling (ABPP), and fluorescence-activated cell sorting (FACS). The probe was able to detect changes in FAO activity induced by chemical modulators in cultured cells. The probe was further employed for fluorescence imaging of FAO in mouse liver tissues and revealed the metabolic heterogeneity of FAO activity in hepatocytes by the combination of FACS and gene expression analysis, highlighting the utility of our probe as a chemical tool for fatty acid metabolism research.

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Triglyceride cycling enables modification of stored fatty acids

et al. 2023

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Triglyceride cycling is the process of continuous degradation and re-synthesis of triglyceride in cellular stores. We show in 3T3-L1 adipocytes that triglycerides are subject to rapid turnover and re-arrangement of fatty acids with an estimated half-life of 2–4 h. We develop a tracing technology that can simultaneously and quantitatively follow the metabolism of multiple fatty acids to study the triglyceride futile substrate cycle directly and with molecular species resolution. Our approach is based on alkyne fatty acid tracers and mass spectrometry. The triglyceride cycling is connected to modification of released fatty acids by elongation and desaturation. Through cycling and modification, saturated fatty acids are slowly converted to monounsaturated fatty acids, and linoleic acid to arachidonic acid. We conclude that triglyceride cycling renders stored fatty acids accessible for metabolic alteration. The overall process facilitates cellular adjustments to the stored fatty acid pool to meet changing needs of the cell.

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Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates

Cited by 6 publications

References 49 publications

Tracing Lipid Metabolism by Alkyne Lipids and Mass Spectrometry: The State of the Art

Tracing Lipid Metabolism by Alkyne Lipids and Mass Spectrometry: The State of the Art

Fluorescence-Based Detection of Fatty Acid β-Oxidation in Cells and Tissues Using Quinone Methide-Releasing Probes

Triglyceride cycling enables modification of stored fatty acids

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