Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates
Fatty acid beta-oxidation is a key process in mammalian lipid catabolism. Disturbance of this process results in severe clinical symptoms, including dysfunction of the liver, a major beta-oxidizing tissue. For a thorough understanding of this process, a comprehensive analysis of involved fatty acid...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-04-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227522000219 |
| _version_ | 1817984127189647360 |
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| author | Lars Kuerschner Philipp Leyendecker Kristina Klizaite Maria Fiedler Jennifer Saam Christoph Thiele |
| author_facet | Lars Kuerschner Philipp Leyendecker Kristina Klizaite Maria Fiedler Jennifer Saam Christoph Thiele |
| author_sort | Lars Kuerschner |
| collection | DOAJ |
| description | Fatty acid beta-oxidation is a key process in mammalian lipid catabolism. Disturbance of this process results in severe clinical symptoms, including dysfunction of the liver, a major beta-oxidizing tissue. For a thorough understanding of this process, a comprehensive analysis of involved fatty acid and acyl-carnitine intermediates is desired, but capable methods are lacking. Here, we introduce oxaalkyne and alkyne fatty acids as novel tracers to study the beta-oxidation of long- and medium-chain fatty acids in liver lysates and primary hepatocytes. Combining these new tracer tools with highly sensitive chromatography and mass spectrometry analyses, this study confirms differences in metabolic handling of fatty acids of different chain length. Unlike longer chains, we found that medium-chain fatty acids that were activated inside or outside of mitochondria by different acyl-CoA synthetases could enter mitochondria in the form of free fatty acids or as carnitine esters. Upon mitochondrial beta-oxidation, shortened acyl-carnitine metabolites were then produced and released from mitochondria. In addition, we show that hepatocytes ultimately also secreted these shortened acyl chains into their surroundings. Furthermore, when mitochondrial beta-oxidation was hindered, we show that peroxisomal beta-oxidation likely acts as a salvage pathway, thereby maintaining the levels of shortened fatty acid secretion. Taken together, we conclude that this new method based on oxaalkyne and alkyne fatty acids allows for metabolic tracing of the beta-oxidation pathway in tissue lysate and in living cells with unique coverage of metabolic intermediates and at unprecedented detail. |
| first_indexed | 2024-04-13T23:40:45Z |
| format | Article |
| id | doaj.art-5b605ae5d5d04288b3c8fc2dd325205c |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-04-13T23:40:45Z |
| publishDate | 2022-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-5b605ae5d5d04288b3c8fc2dd325205c2022-12-22T02:24:30ZengElsevierJournal of Lipid Research0022-22752022-04-01634100188Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediatesLars Kuerschner0Philipp Leyendecker1Kristina Klizaite2Maria Fiedler3Jennifer Saam4Christoph Thiele5For correspondence: Lars Kuerschner; LIMES Life and Medical Sciences Institute, University of Bonn, Bonn, GermanyLIMES Life and Medical Sciences Institute, University of Bonn, Bonn, GermanyLIMES Life and Medical Sciences Institute, University of Bonn, Bonn, GermanyLIMES Life and Medical Sciences Institute, University of Bonn, Bonn, GermanyLIMES Life and Medical Sciences Institute, University of Bonn, Bonn, GermanyLIMES Life and Medical Sciences Institute, University of Bonn, Bonn, GermanyFatty acid beta-oxidation is a key process in mammalian lipid catabolism. Disturbance of this process results in severe clinical symptoms, including dysfunction of the liver, a major beta-oxidizing tissue. For a thorough understanding of this process, a comprehensive analysis of involved fatty acid and acyl-carnitine intermediates is desired, but capable methods are lacking. Here, we introduce oxaalkyne and alkyne fatty acids as novel tracers to study the beta-oxidation of long- and medium-chain fatty acids in liver lysates and primary hepatocytes. Combining these new tracer tools with highly sensitive chromatography and mass spectrometry analyses, this study confirms differences in metabolic handling of fatty acids of different chain length. Unlike longer chains, we found that medium-chain fatty acids that were activated inside or outside of mitochondria by different acyl-CoA synthetases could enter mitochondria in the form of free fatty acids or as carnitine esters. Upon mitochondrial beta-oxidation, shortened acyl-carnitine metabolites were then produced and released from mitochondria. In addition, we show that hepatocytes ultimately also secreted these shortened acyl chains into their surroundings. Furthermore, when mitochondrial beta-oxidation was hindered, we show that peroxisomal beta-oxidation likely acts as a salvage pathway, thereby maintaining the levels of shortened fatty acid secretion. Taken together, we conclude that this new method based on oxaalkyne and alkyne fatty acids allows for metabolic tracing of the beta-oxidation pathway in tissue lysate and in living cells with unique coverage of metabolic intermediates and at unprecedented detail.http://www.sciencedirect.com/science/article/pii/S0022227522000219clicklipid tracerβ-oxidationmid-chain fatty acidCPTfatty acid metabolism |
| spellingShingle | Lars Kuerschner Philipp Leyendecker Kristina Klizaite Maria Fiedler Jennifer Saam Christoph Thiele Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates Journal of Lipid Research click lipid tracer β-oxidation mid-chain fatty acid CPT fatty acid metabolism |
| title | Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates |
| title_full | Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates |
| title_fullStr | Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates |
| title_full_unstemmed | Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates |
| title_short | Development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta-oxidation pathways and intermediates |
| title_sort | development of oxaalkyne and alkyne fatty acids as novel tracers to study fatty acid beta oxidation pathways and intermediates |
| topic | click lipid tracer β-oxidation mid-chain fatty acid CPT fatty acid metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S0022227522000219 |
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