1,519 results on '"acyl-coA"'
Search Results
2. Metabolic engineering of Corynebacterium glutamicum for L-alanine production
- Author
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Huang, Yu, Li, Hedan, Zhao, Guihong, Hu, Xiaoqing, and Wang, Xiaoyuan
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- 2024
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3. Functions of Coenzyme A and Acyl-CoA in Post-Translational Modification and Human Disease
- Author
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Jumin Xie, Zhang Yu, Ying Zhu, Mei Zheng, and Yanfang Zhu
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acylation ,acyl-coa ,cancer ,coenzyme a ,post-translational modification ,Biochemistry ,QD415-436 ,Biology (General) ,QH301-705.5 - Abstract
Coenzyme A (CoA) is synthesized from pantothenate, L-cysteine and adenosine triphosphate (ATP), and plays a vital role in diverse physiological processes. Protein acylation is a common post-translational modification (PTM) that modifies protein structure, function and interactions. It occurs via the transfer of acyl groups from acyl-CoAs to various amino acids by acyltransferase. The characteristics and effects of acylation vary according to the origin, structure, and location of the acyl group. Acetyl-CoA, formyl-CoA, lactoyl-CoA, and malonyl-CoA are typical acyl group donors. The major acyl donor, acyl-CoA, enables modifications that impart distinct biological functions to both histone and non-histone proteins. These modifications are crucial for regulating gene expression, organizing chromatin, managing metabolism, and modulating the immune response. Moreover, CoA and acyl-CoA play significant roles in the development and progression of neurodegenerative diseases, cancer, cardiovascular diseases, and other health conditions. The goal of this review was to systematically describe the types of commonly utilized acyl-CoAs, their functions in protein PTM, and their roles in the progression of human diseases.
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- 2024
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4. 代谢工程改造酿酒酵母高效合成游离脂肪酸.
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朱满志, 陈献忠, 沈微, 杨海泉, and 夏媛媛
- Abstract
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- 2024
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5. Acetyl-CoA synthetase (ACSS2) does not generate butyryl- and crotonyl-CoA
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Nour Zeaiter, Laura Belot, Valérie Cunin, Roland Abi Nahed, Malgorzata Tokarska-Schlattner, Audrey Le Gouellec, Carlo Petosa, Saadi Khochbin, and Uwe Schlattner
- Subjects
Acyl-CoA ,Epigenetics ,Histone acylation ,Protein acylation ,Substrate specificity ,Internal medicine ,RC31-1245 - Abstract
Acetyl and other acyl groups from different short-chain fatty acids (SCFA) competitively modify histones at various lysine sites. To fully understand the functional significance of such histone acylation, a key epigenetic mechanism, it is crucial to characterize the cellular sources of the corresponding acyl-CoA molecules required for the lysine modification. Like acetate, SCFAs such as propionate, butyrate and crotonate are thought to be the substrates used to generate the corresponding acyl-CoAs by enzymes known as acyl-CoA synthetases. The acetyl-CoA synthetase, ACSS2, which produces acetyl-CoA from acetate in the nucleocytoplasmic compartment, has been proposed to also mediate the synthesis of acyl-CoAs such as butyryl- and crotonyl-CoA from the corresponding SCFAs. This idea is now widely accepted and is sparking new research projects. However, based on our direct in vitro experiments with purified or recombinant enzymes and structural considerations, we demonstrate that ACSS2 is unable to mediate the generation of non-acetyl acyl-CoAs like butyryl- and crotonyl-CoA. It is therefore essential to re-examine published data and corresponding discussions in the light of this new finding.
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- 2024
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6. Multi-Method Quantification of Acetyl-Coenzyme A and Further Acyl-Coenzyme A Species in Normal and Ischemic Rat Liver.
- Author
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Tokarska-Schlattner, Malgorzata, Zeaiter, Nour, Cunin, Valérie, Attia, Stéphane, Meunier, Cécile, Kay, Laurence, Achouri, Amel, Hiriart-Bryant, Edwige, Couturier, Karine, Tellier, Cindy, El Harras, Abderrafek, Elena-Herrmann, Bénédicte, Khochbin, Saadi, Le Gouellec, Audrey, and Schlattner, Uwe
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ACETYLCOENZYME A , *KREBS cycle , *ACYL coenzyme A , *LIVER , *RATS - Abstract
Thioesters of coenzyme A (CoA) carrying different acyl chains (acyl-CoAs) are central intermediates of many metabolic pathways and donor molecules for protein lysine acylation. Acyl-CoA species largely differ in terms of cellular concentrations and physico-chemical properties, rendering their analysis challenging. Here, we compare several approaches to quantify cellular acyl-CoA concentrations in normal and ischemic rat liver, using HPLC and LC-MS/MS for multi-acyl-CoA analysis, as well as NMR, fluorimetric and spectrophotometric techniques for the quantification of acetyl-CoAs. In particular, we describe a simple LC-MS/MS protocol that is suitable for the relative quantification of short and medium-chain acyl-CoA species. We show that ischemia induces specific changes in the short-chain acyl-CoA relative concentrations, while mild ischemia (1–2 min), although reducing succinyl-CoA, has little effects on acetyl-CoA, and even increases some acyl-CoA species upstream of the tricarboxylic acid cycle. In contrast, advanced ischemia (5–6 min) also reduces acetyl-CoA levels. Our approach provides the keys to accessing the acyl-CoA metabolome for a more in-depth analysis of metabolism, protein acylation and epigenetics. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
7. The Physiological and Pathological Role of Acyl-CoA Oxidation.
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Szrok-Jurga, Sylwia, Czumaj, Aleksandra, Turyn, Jacek, Hebanowska, Areta, Swierczynski, Julian, Sledzinski, Tomasz, and Stelmanska, Ewa
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FATTY acid oxidation , *ACYL coenzyme A , *ALIMENTARY canal , *GASTROINTESTINAL system , *LEUCOCYTES , *HEART , *LUNGS - Abstract
Fatty acid metabolism, including β-oxidation (βOX), plays an important role in human physiology and pathology. βOX is an essential process in the energy metabolism of most human cells. Moreover, βOX is also the source of acetyl-CoA, the substrate for (a) ketone bodies synthesis, (b) cholesterol synthesis, (c) phase II detoxication, (d) protein acetylation, and (d) the synthesis of many other compounds, including N-acetylglutamate—an important regulator of urea synthesis. This review describes the current knowledge on the importance of the mitochondrial and peroxisomal βOX in various organs, including the liver, heart, kidney, lung, gastrointestinal tract, peripheral white blood cells, and other cells. In addition, the diseases associated with a disturbance of fatty acid oxidation (FAO) in the liver, heart, kidney, lung, alimentary tract, and other organs or cells are presented. Special attention was paid to abnormalities of FAO in cancer cells and the diseases caused by mutations in gene-encoding enzymes involved in FAO. Finally, issues related to α- and ω- fatty acid oxidation are discussed. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
8. Brain CoA and Acetyl CoA Metabolism in Mechanisms of Neurodegeneration.
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Moiseenok, Andrey G. and Kanunnikova, Nina P.
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NEURAL transmission , *ACETYLCOENZYME A , *TAU proteins , *BLOOD-brain barrier , *PANTOTHENIC acid , *NEURODEGENERATION , *ALZHEIMER'S disease , *PARKINSON'S disease - Abstract
The processes of biotransformation of pantothenic acid (Pan) in the biosynthesis and hydrolysis of CoA, key role of pantothenate kinase (PANK) and CoA synthetase (CoASY) in the formation of the priority mitochondrial pool of CoA, with a high metabolic turnover of the coenzyme and limited transport of Pan across the blood-brain barrier are considered. The system of acetyl-CoA, a secondary messenger, which is the main substrate of acetylation processes including formation of N-acetyl aspartate and acetylcholine, post-translational modification of histones, predetermines protection of the neurons against degenerative signals and cholinergic neurotransmission. Biochemical mechanisms of neurodegenerative syndromes in the cases of PANK and CoASY defects, and the possibility of correcting of CoA biosynthesis in the models with knockouts of these enzymes have been described. The data of a post-mortem study of the brains from the patients with Huntington's and Alzheimer's diseases are presented, proving Pan deficiency in the CNS, which is especially pronounced in the pathognomonic neurostructures. In the frontal cortex of the patients with Parkinson's disease, combined immunofluorescence of anti-CoA- and anti-tau protein was detected, reflecting CoAlation during dimerization of the tau protein and its redox sensitivity. Redox activity and antioxidant properties of the precursors of CoA biosynthesis were confirmed in vitro with synaptosomal membranes and mitochondria during modeling of aluminum neurotoxicity accompanied by the decrease in the level of CoA in CNS. The ability of CoA biosynthesis precursors to stabilize glutathione pool in neurostructures, in particular, in the hippocampus, is considered as a pathogenetic protection mechanism during exposure to neurotoxins, development of neuroinflammation and neurodegeneration, and justifies the combined use of Pan derivatives (for example, D-panthenol) and glutathione precursors (N-acetylcysteine). Taking into account the discovery of new functions of CoA (redox-dependent processes of CoAlation of proteins, possible association of oxidative stress and deficiency of Pan (CoA) in neurodegenerative pathology), it seems promising to study bioavailability and biotransformation of Pan derivatives, in particular of D-panthenol, 4′-phospho-pantetheine, its acylated derivatives, and compositions with redox pharmacological compounds, are promising for their potential use as etiopathogenetic agents. [ABSTRACT FROM AUTHOR]
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- 2023
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9. Assessing the biotechnological potential of cotton type-1 and type-2 diacylglycerol acyltransferases in transgenic systems.
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Shockey, Jay, Parchuri, Prasad, Thyssen, Gregory N., and Bates, Philip D.
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ACYLTRANSFERASES , *VEGETABLE oils , *TRANSGENIC seeds , *COTTON , *TRANSGENIC plants , *OILSEEDS , *ISOENZYMES - Abstract
The chemical and physical properties of vegetable oils are largely dictated by the ratios of 4–6 common fatty acids contained within each oil. However, examples of plant species that accumulate from trace amounts to >90% of certain unusual fatty acids in seed triacylglycerols have been reported. Many of the general enzymatic reactions that drive both common and unusual fatty acid biosynthesis and accumulation in stored lipids are known, but which isozymes have evolved to specifically fill this role and how they coordinate in vivo is still poorly understood. Cotton (Gossypium sp.) is the very rare example of a commodity oilseed that produces biologically relevant amounts of unusual fatty acids in its seeds and other organs. In this case, unusual cyclopropyl fatty acids (named after the cyclopropane and cyclopropene moieties within the fatty acids) are found in membrane and storage glycerolipids (e.g. seed oils). Such fatty acids are useful in the synthesis of lubricants, coatings, and other types of valuable industrial feedstocks. To characterize the role of cotton acyltransferases in cyclopropyl fatty acid accumulation for bioengineering applications, we cloned and characterized type-1 and type-2 diacylglycerol acyltransferases from cotton and compared their biochemical properties to that of litchi (Litchi chinensis), another cyclopropyl fatty acid-producing plant. The results presented from transgenic microbes and plants indicate both cotton DGAT1 and DGAT2 isozymes efficiently utilize cyclopropyl fatty acid-containing substrates, which helps to alleviate biosynthetic bottlenecks and enhances total cyclopropyl fatty acid accumulation in the seed oil. • Cotton contains two DGAT1 genes and ten DGAT2 orthologs. • Metabolic bottlenecks typically limit CPFA production in transgenic seeds. • Cotton and litchi DGATs alleviate the bottleneck and enhance CPFA accumulation in transgenic yeast and plants. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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10. LsSpt23p is a regulator of triacylglycerol synthesis in the oleaginous yeast Lipomyces starkeyi.
- Author
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Takaku, Hiroaki, Kazama, Haruka, Sato, Rikako, Mori, Kazuki, Ara, Satoshi, Ishiya, Koji, Matsuzawa, Tomohiko, Yaoi, Katsuro, Araki, Hideo, Shida, Yosuke, Ogasawara, Wataru, Tashiro, Kosuke, Kuhara, Satoru, Yamazaki, Harutake, and Aburatani, Sachiyo
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FATTY acid desaturase , *GENE expression , *YEAST , *BIOSYNTHESIS , *PHOSPHATIDIC acids , *CITRATES , *PHOSPHOCHOLINE - Abstract
The oleaginous yeast Lipomyces starkeyi has considerable potential in industrial application, since it can accumulate a large amount of triacylglycerol (TAG), which is produced from sugars under nitrogen limitation condition. However, the regulation of lipogenesis in L. starkeyi has not been investigated in depth. In this study, we compared the genome sequences of wild-type and mutants with increased TAG productivity, and identified a regulatory protein, LsSpt23p, which contributes to the regulation of TAG synthesis in L. starkeyi. L. starkeyi mutants overexpressing LsSPT23 had increased TAG productivity compared with the wild-type strain. Quantitative real-time PCR analysis showed that LsSpt23p upregulated the expression of GPD1, which encodes glycerol 3-phosphate dehydrogenase; the Kennedy pathway genes SCT1, SLC1, PAH1, DGA1, and DGA2; the citrate-mediated acyl-CoA synthesis pathway-related genes ACL1, ACL2, ACC1, FAS1, and FAS2; and OLE1, which encodes ∆9 fatty acid desaturase. Chromatin immunoprecipitation-quantitative PCR assays indicated that LsSpt23p acts as a direct regulator of SLC1 and PAH1, all the citrate-mediated acyl-CoA synthesis pathway–related genes, and OLE1. These results indicate that LsSpt23p regulates TAG synthesis. Phosphatidic acid is a common substrate of phosphatidic acid phosphohydrolase, which is used for TAG synthesis, and phosphatidate cytidylyltransferase 1 for phospholipid synthesis in the Kennedy pathway. LsSpt23p directly regulated PAH1 but did not affect the expression of CDS1, suggesting that the preferred route of carbon is the Pah1p-mediated TAG synthesis pathway under nitrogen limitation condition. The present study contributes to understanding the regulation of TAG synthesis, and will be valuable in future improvement of TAG productivity in oleaginous yeasts. Key points: LsSpt23p was identified as a positive regulator of TAG biosynthesis LsSPT23 overexpression enhanced TAG biosynthesis gene expression and TAG production LsSPT23M1108Toverexpression mutant showed fivefold higher TAG production than control [ABSTRACT FROM AUTHOR]
- Published
- 2023
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- View/download PDF
11. Ischemia promotes acyl-CoAs dephosphorylation and propionyl-CoA accumulation.
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He, Wentao, Berthiaume, Jessica M., Previs, Stephen, Kasumov, Takhar, and Zhang, Guo-Fang
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DEPHOSPHORYLATION , *ISCHEMIA , *HEART injuries , *ACYL coenzyme A , *MYOCARDIAL ischemia , *PHOSPHOCREATINE - Abstract
Introduction: Our untargeted metabolic data unveiled that Acyl-CoAs undergo dephosphorylation, however little is known about these novel metabolites and their physiology/pathology relevance. Objectives: To understand the relationship between acyl-CoAs dephosphorylation and energy status as implied in our previous work, we seek to investigate how ischemia (energy depletion) triggers metabolic changes, specifically acyl-CoAs dephosphorylation in this work. Methods: Rat hearts were isolated and perfused in Langendorff mode for 15 min followed by 0, 5, 15, and 30 minutes of global ischemia. The heart tissues were harvested for metabolic analysis. Results: As expected, ATP and phosphocreatine were significantly decreased during ischemia. Most short- and medium-chain acyl-CoAs progressively increased with ischemic time from 0 to 15 min, whereas a 30-minute ischemia did not lead to further change. Unlike other acyl-CoAs, propionyl-CoA accumulated progressively in the hearts that underwent ischemia from 0 to 30 min. Progressive dephosphorylation occurred to all assayed acyl-CoAs and free CoA regardless their level changes during the ischemia. Conclusion: The present work further confirms that dephosphorylation of acyl-CoAs is an energy-dependent process and how this dephosphorylation is mediated warrants further investigations. It is plausible that dephosphorylation of acyl-CoAs and limited anaplerosis are involved in ischemic injuries to heart. Further investigations are warranted to examine the mechanisms of acyl-CoA dephosphorylation and how the dephosphorylation is possibly involved in ischemic injuries. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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12. Protein acylation links metabolism and the control of signal transduction, transcription regulation, growth, and pathogenicity in Actinobacteria.
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Peng, Zhi‐Yao, Fu, Yu, Zhao, Liu‐Chang, Dong, Yu‐Qi, Chen, Zong‐Qin, You, Di, and Ye, Bang‐Ce
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METABOLIC regulation , *GENETIC transcription regulation , *CELLULAR signal transduction , *ACYLATION , *ACTINOBACTERIA , *SOMATIC cell nuclear transfer - Abstract
Actinobacteria have a complex life cycle, including morphological and physiological differentiation which are often associated with the biosynthesis of secondary metabolites. Recently, increased interest in post‐translational modifications (PTMs) in these Gram‐positive bacteria has highlighted the importance of PTMs as signals that provide functional diversity and regulation by modifying proteins to respond to diverse stimuli. Here, we review the developments in research on acylation, a typical PTM that uses acyl‐CoA or related metabolites as donors, as well as the understanding of the direct link provided by acylation between cell metabolism and signal transduction, transcriptional regulation, cell growth, and pathogenicity in Actinobacteria. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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13. Defects in lipid homeostasis reflect the function of TANGO2 in phospholipid and neutral lipid metabolism
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Agustin Leonardo Lujan, Ombretta Foresti, Conor Sugden, Nathalie Brouwers, Alex Mateo Farre, Alessio Vignoli, Mahshid Azamian, Alicia Turner, Jose Wojnacki, and Vivek Malhotra
- Subjects
TANGO2 ,lipid droplet ,reactive oxygen species ,acyl-CoA ,lysophosphatidic acid ,lipid peroxidation ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
We show that TANGO2 in mammalian cells localizes predominantly to mitochondria and partially at mitochondria sites juxtaposed to lipid droplets (LDs) and the endoplasmic reticulum. HepG2 cells and fibroblasts of patients lacking TANGO2 exhibit enlarged LDs. Quantitative lipidomics revealed a marked increase in lysophosphatidic acid (LPA) and a concomitant decrease in its biosynthetic precursor phosphatidic acid (PA). These changes were exacerbated in nutrient-starved cells. Based on our data, we suggest that TANGO2 function is linked to acyl-CoA metabolism, which is necessary for the acylation of LPA to generate PA. The defect in acyl-CoA availability impacts the metabolism of many other fatty acids, generates high levels of reactive oxygen species, and promotes lipid peroxidation. We suggest that the increased size of LDs is a combination of enrichment in peroxidized lipids and a defect in their catabolism. Our findings help explain the physiological consequence of mutations in TANGO2 that induce acute metabolic crises, including rhabdomyolysis, cardiomyopathy, and cardiac arrhythmias, often leading to fatality upon starvation and stress.
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- 2023
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14. Time‐resolved multiparameter analytics on a cell‐free production platform for acyl‐CoA precursors.
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Maehler, Dominic, Hoefgen, Sandra, Münchberg, Ute, Schmitz, Oliver J., Rautschek, Julia, Huang, Ying, Freier, Erik, and Valiante, Vito
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ACYL coenzyme A ,CAPILLARY electrophoresis ,ENZYMATIC analysis ,ACETYLCOENZYME A ,BIOSYNTHESIS ,LEAD time (Supply chain management) - Abstract
Cell‐free biosynthesis is emerging as a very attractive alternative for the production of market‐relevant molecules. The free combination of enzymes, regardless of where they are isolated from, raises the possibility to build more efficient synthetic routes but at the same time leads to higher complexity regarding the analysis of the different enzymatic steps. Here we present an analytical method for the real‐time analysis of acyl‐CoA blocks forming and consuming during multi‐step catalyses. We focused on malonyl‐Coenzyme A and acetyl‐CoA, which are the most used acyl‐CoA units for carbon chain elongations. By employing capillary electrophoresis, we could detect the decrease of educts and the formation of products in a time‐resolved fashion. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
15. Suppression of RCAN1 alleviated lipid accumulation and mitochondrial fission in diabetic cardiomyopathy.
- Author
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Shu, Songren, Cui, Hao, Liu, Zirui, Zhang, Hang, Yang, Yicheng, Chen, Xiao, Zeng, Zhiwei, Du, Leilei, Fu, Mengxia, Yang, Ziang, Wang, Peizhi, Wang, Chuangshi, Gao, Huimin, Yang, Qiaoxi, Lin, Xiaojun, Yang, Tianshuo, Chen, Zhice, Wu, Sijin, Wang, Xiaohu, and Zhao, Ruojin
- Subjects
MITOCHONDRIAL dynamics ,DIABETIC cardiomyopathy ,HEART diseases ,HEART failure ,HEART transplantation - Abstract
Although metabolic disturbance is a characteristic of diabetic cardiomyopathy (DbCM), the detailed pathogenesis of DbCM remains unknown. We used a heart transplantation (HTx) cohort to explore the effect of diabetes mellitus on heart failure (HF) progression dependent of myocardium. Microscopic and ultramicroscopic pathology were used to depict the pathological features of human myocardium of DbCM. We performed targeted metabolomics to characterize the metabolic phenotype of human DbCM. Transcriptomics data were analyzed and weighted gene co-expression network analysis was performed to explore the potential upstream regulator for metabolic remodeling of DbCM. In vivo and in vitro experiments were further conducted to demonstrate the therapeutic effects and molecular mechanisms. DbCM promoted the progression of HF and increased death or HF-rehospitalization after HTx. Lipid accumulation and mitochondrial fission were the obvious pathological features of DbCM myocardium. The concentrations of C14:0-CoA and C16:1-CoA were significantly increased in the myocardium, and they were positively correlated with the accelerated HF progression and RCAN1 expression in DbCM patients. Knockdown of RCAN1 improved cardiac dysfunction, lipid accumulation, and mitochondrial fission in db/db mice. In vitro studies showed that RCAN1 knockdown improved mitochondrial dysfunction in DbCM cardiomyocytes via the RCAN1-p-Drp1 Ser
616 axis. Diabetes is associated with faster progression of HF and causes poor prognosis after HTx, accompanied by metabolic remodeling in the myocardium. Accumulation of long chain acyl-CoA in the myocardium is the metabolic hallmark of human DbCM and is associated with more rapid disease progression for DbCM patients. Upregulation of RCAN1 in the myocardium is associated with the metabolic signatures of DbCM and RCAN1 is a potential therapeutic target for DbCM. [Display omitted] • Diabetes promotes non-diabetic heart failure after heart transplantation. • The increase in acyl-CoA is the metabolic hallmark of diabetic cardiomyopathy. • Knockdown of RCAN1 improved the cardiac function and structure of db/db mice. • RCAN1-p-Drp1 Ser616 axis contributes to mitochondrial dysfunction in cardiomyocytes. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
16. The Physiological and Pathological Role of Acyl-CoA Oxidation
- Author
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Sylwia Szrok-Jurga, Aleksandra Czumaj, Jacek Turyn, Areta Hebanowska, Julian Swierczynski, Tomasz Sledzinski, and Ewa Stelmanska
- Subjects
beta-oxidation ,peroxisomal fatty acid oxidation ,acyl-CoA ,fatty acid metabolism ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Fatty acid metabolism, including β-oxidation (βOX), plays an important role in human physiology and pathology. βOX is an essential process in the energy metabolism of most human cells. Moreover, βOX is also the source of acetyl-CoA, the substrate for (a) ketone bodies synthesis, (b) cholesterol synthesis, (c) phase II detoxication, (d) protein acetylation, and (d) the synthesis of many other compounds, including N-acetylglutamate—an important regulator of urea synthesis. This review describes the current knowledge on the importance of the mitochondrial and peroxisomal βOX in various organs, including the liver, heart, kidney, lung, gastrointestinal tract, peripheral white blood cells, and other cells. In addition, the diseases associated with a disturbance of fatty acid oxidation (FAO) in the liver, heart, kidney, lung, alimentary tract, and other organs or cells are presented. Special attention was paid to abnormalities of FAO in cancer cells and the diseases caused by mutations in gene-encoding enzymes involved in FAO. Finally, issues related to α- and ω- fatty acid oxidation are discussed.
- Published
- 2023
- Full Text
- View/download PDF
17. Multi-Method Quantification of Acetyl-Coenzyme A and Further Acyl-Coenzyme A Species in Normal and Ischemic Rat Liver
- Author
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Malgorzata Tokarska-Schlattner, Nour Zeaiter, Valérie Cunin, Stéphane Attia, Cécile Meunier, Laurence Kay, Amel Achouri, Edwige Hiriart-Bryant, Karine Couturier, Cindy Tellier, Abderrafek El Harras, Bénédicte Elena-Herrmann, Saadi Khochbin, Audrey Le Gouellec, and Uwe Schlattner
- Subjects
acetyl-CoA ,acyl-CoA ,fluorometric assay ,HPLC ,liver ischemia ,mass spectrometry ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Thioesters of coenzyme A (CoA) carrying different acyl chains (acyl-CoAs) are central intermediates of many metabolic pathways and donor molecules for protein lysine acylation. Acyl-CoA species largely differ in terms of cellular concentrations and physico-chemical properties, rendering their analysis challenging. Here, we compare several approaches to quantify cellular acyl-CoA concentrations in normal and ischemic rat liver, using HPLC and LC-MS/MS for multi-acyl-CoA analysis, as well as NMR, fluorimetric and spectrophotometric techniques for the quantification of acetyl-CoAs. In particular, we describe a simple LC-MS/MS protocol that is suitable for the relative quantification of short and medium-chain acyl-CoA species. We show that ischemia induces specific changes in the short-chain acyl-CoA relative concentrations, while mild ischemia (1–2 min), although reducing succinyl-CoA, has little effects on acetyl-CoA, and even increases some acyl-CoA species upstream of the tricarboxylic acid cycle. In contrast, advanced ischemia (5–6 min) also reduces acetyl-CoA levels. Our approach provides the keys to accessing the acyl-CoA metabolome for a more in-depth analysis of metabolism, protein acylation and epigenetics.
- Published
- 2023
- Full Text
- View/download PDF
18. Does chromatin function as a metabolite reservoir?
- Author
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Nirello, Vinícius D., Rodrigues de Paula, Dieggo, Araújo, Nathália V.P., and Varga-Weisz, Patrick D.
- Subjects
- *
CHROMATIN , *POST-translational modification , *ACYL coenzyme A , *GENE expression , *ACYLATION - Abstract
Alternative histone acylations integrate gene expression with cellular metabolic states. Recent measurements of cellular acyl-coenzyme A (acyl-CoA) pools highlight the potential that histone post-translational modifications (PTMs) contribute directly to the regulation of metabolite pools. A metabolite-centric view throws new light onto roles and evolution of histone PTMs. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
19. Functions of Coenzyme A and Acyl-CoA in Post-Translational Modification and Human Disease.
- Author
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Xie J, Yu Z, Zhu Y, Zheng M, and Zhu Y
- Subjects
- Humans, Neurodegenerative Diseases metabolism, Neurodegenerative Diseases genetics, Neoplasms metabolism, Neoplasms genetics, Acylation, Cardiovascular Diseases metabolism, Cardiovascular Diseases genetics, Animals, Protein Processing, Post-Translational, Acyl Coenzyme A metabolism, Coenzyme A metabolism
- Abstract
Coenzyme A (CoA) is synthesized from pantothenate, L-cysteine and adenosine triphosphate (ATP), and plays a vital role in diverse physiological processes. Protein acylation is a common post-translational modification (PTM) that modifies protein structure, function and interactions. It occurs via the transfer of acyl groups from acyl-CoAs to various amino acids by acyltransferase. The characteristics and effects of acylation vary according to the origin, structure, and location of the acyl group. Acetyl-CoA, formyl-CoA, lactoyl-CoA, and malonyl-CoA are typical acyl group donors. The major acyl donor, acyl-CoA, enables modifications that impart distinct biological functions to both histone and non-histone proteins. These modifications are crucial for regulating gene expression, organizing chromatin, managing metabolism, and modulating the immune response. Moreover, CoA and acyl-CoA play significant roles in the development and progression of neurodegenerative diseases, cancer, cardiovascular diseases, and other health conditions. The goal of this review was to systematically describe the types of commonly utilized acyl-CoAs, their functions in protein PTM, and their roles in the progression of human diseases., (© 2024 The Author(s). Published by IMR Press.)
- Published
- 2024
- Full Text
- View/download PDF
20. Substrate Specificity of the 3-Methylmercaptopropionyl Coenzyme A Dehydrogenase (DmdC1) from Ruegeria pomeroyi DSS-3.
- Author
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Tao Wang, Hao Shi, and Whitman, William B.
- Subjects
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FATTY acid oxidation , *MARINE bacteria , *ACYL coenzyme A , *DEMETHYLATION , *DEHYDROGENASES , *ACYLTRANSFERASES - Abstract
The acyl-coenzyme A (CoA) dehydrogenase family enzyme DmdC catalyzes the third step in the dimethylsulfoniopropionate (DMSP) demethylation pathway, the oxidation of 3-methylmercaptopropionyl-CoA (MMPA-CoA) to 3-methylthioacryloyl-CoA (MTA-CoA). To study its substrate specificity, the recombinant DmdC1 from Ruegeria pomeroyi was characterized. In addition to MMPA-CoA, the enzyme was highly active with short-chain acyl-CoAs, with Km values for MMPA-CoA, butyryl-CoA, valeryl-CoA, caproyl-CoA, heptanoyl-CoA, caprylyl-CoA, and isobutyryl-CoA of 36, 19, 7, 11, 14, 10, and 149 μM, respectively, and kcat values of 1.48, 0.40, 0.48, 0.73, 0.46, 0.23, and 0.01 s-1, respectively. Among these compounds, MMPA-CoA was the best substrate. The high affinity of DmdC1 for its substrate supports the model for kinetic regulation of the demethylation pathway. In contrast to DmdB, which catalyzes the formation of MMPA-CoA from MMPA, CoA, and ATP, DmdC1 was not affected by physiological concentrations of potential effectors, such as DMSP, MMPA, ATP, and ADP. Thus, compared to the other enzymes of the DMSP demethylation pathway, DmdC1 has only minimal adaptations for DMSP metabolism compared to other enzymes in the same family with similar substrates, supporting the hypothesis that it evolved relatively recently from a short-chain acyl-CoA dehydrogenase involved in fatty acid oxidation. IMPORTANCE We report the kinetic properties of DmdC1 from the model organism R. pomeroyi and close an important gap in the literature. While the crystal structure of this enzyme was recently solved and its mechanism of action described (X. Shao, H. Y. Cao, F. Zhao, M. Peng, et al., Mol Microbiol 111:1057-1073, 2019, https://doi.org/10 .1111/mmi.14211), its substrate specificity and sensitivity to potential effectors was never examined. We show that DmdC1 has a high affinity for other short-chain acylCoAs in addition to MMPA-CoA, which is the natural substrate in DMSP metabolism and is not affected by the potential effectors tested. This evidence supports the hypothesis that DmdC1 possesses few adaptations to DMSP metabolism and likely evolved relatively recently from a short-chain acyl-CoA dehydrogenase involved in fatty acid oxidation. This work is important because it expands our understanding of the adaptation of marine bacteria to the increased availability of DMSP about 250 million years ago. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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21. p-Nitrophenyl esters provide new insights and applications for the thiolase enzyme OleA
- Author
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Megan D. Smith, Lambros J. Tassoulas, Troy A. Biernath, Jack E. Richman, Kelly G. Aukema, and Lawrence P. Wackett
- Subjects
OleA ,Thiolase ,Claisen condensation ,Bacteria ,p-Nitrophenyl ester ,Acyl-CoA ,Biotechnology ,TP248.13-248.65 - Abstract
The OleA enzyme is distinct amongst thiolase enzymes in binding two long (≥C8) acyl chains into structurally-opposed hydrophobic channels, denoted A and B, to carry out a non-decarboxylative Claisen condensation reaction and initiate the biosynthesis of membrane hydrocarbons and β-lactone natural products. OleA has now been identified in hundreds of diverse bacteria via bioinformatics and high-throughput screening using p-nitrophenyl alkanoate esters as surrogate substrates. In the present study, p-nitrophenyl esters were used to probe the reaction mechanism of OleA and shown to be incorporated into Claisen condensation products for the first time. p-Nitrophenyl alkanoate substrates alone were shown not to undergo Claisen condensation, but co-incubation of p-nitrophenyl esters and CoA thioesters produced mixed Claisen products. Mixed product reactions were shown to initiate via acyl group transfer from a p-nitrophenyl carrier to the enzyme active site cysteine, C143. Acyl chains esterified to p-nitrophenol were synthesized and shown to undergo Claisen condensation with an acyl-CoA substrate, showing potential to greatly expand the range of possible Claisen products. Using p-nitrophenyl 1-13C-decanoate, the Channel A bound thioester chain was shown to act as the Claisen nucleophile, representing the first direct evidence for the directionality of the Claisen reaction in any OleA enzyme. These results both provide new insights into OleA catalysis and open a path for making unnatural hydrocarbon and β-lactone natural products for biotechnological applications using cheap and easily synthesized p-nitrophenyl esters.
- Published
- 2021
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22. From benzodiazepines to fatty acids and beyond: revisiting the role of ACBP/DBI.
- Author
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Alquier, Thierry, Christian-Hinman, Catherine A., Alfonso, Julieta, and Færgeman, Nils J.
- Subjects
- *
FATTY acids , *CARRIER proteins , *BENZODIAZEPINES , *ACETYL-CoA carboxylase , *GABA receptors , *GLUTAMINE synthetase , *POLYPEPTIDES - Abstract
Four decades ago Costa and colleagues identified a small, secreted polypeptide in the brain that can displace the benzodiazepine diazepam from the GABA A receptor, and was thus termed diazepam binding inhibitor (DBI). Shortly after, an identical polypeptide was identified in liver by its ability to induce termination of fatty acid synthesis, and was named acyl-CoA binding protein (ACBP). Since then, ACBP/DBI has been studied in parallel without a clear and integrated understanding of its dual roles. The first genetic loss-of-function models have revived the field, allowing targeted approaches to better understand the physiological roles of ACBP/DBI in vivo. We discuss the roles of ACBP/DBI in central and tissue-specific functions in mammals, with an emphasis on metabolism and mechanisms of action. DBI was identified on its ability to displace diazepam from the benzodiazepine binding site on GABA A receptors. ACBP was identified on its ability to induce termination of fatty acid synthesis in goat mammary gland. ACBP/DBI binds long-chain fatty acyl-CoA esters (LCACoAs) with very high affinity. DBI/ACBP is secreted by the unconventional secretory pathway and can be cleaved to regulatory signaling peptides including triakontatetraneuropeptide (ACBP/DBI 1 7– 50) and octadecaneuropeptide (ACBP/DBI 3 3– 50). As an extracellular protein, ACBP/DBI and its derived peptides have pleiotropic effects on gut and pancreatic hormone secretion, neurogenesis, neuronal survival, cognition, and behavior, as well as on energy balance neurocircuits. As an intracellular protein, ACBP/DBI sequesters LCACoAs and mediates their flux to enzymes including ceramide synthases. ACBP/DBI relieves LCACoA-mediated inhibition of enzymes such as acetyl-CoA carboxylase and acyl-CoA synthetases. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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23. Modulation of FadR Binding Capacity for Acyl-CoA Fatty Acids Through Structure-Guided Mutagenesis
- Author
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Martí-Arbona, Ricardo [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)]
- Published
- 2015
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24. Tracking synthesis and turnover of triacylglycerol in leaves
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Ohlrogge, John [Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Research Center. Dept. of Plant Biology. Dept. of Energy]
- Published
- 2015
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25. Reconstruction of Secondary Metabolic Pathway to Synthesize Novel Metabolite in Saccharopolyspora erythraea
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Chong-Yang Ren, Yong Liu, Wen-Ping Wei, Junbiao Dai, and Bang-Ce Ye
- Subjects
CRISPR-Cas9 ,Saccharopolyspora erythraea ,polyketide ,Acyl-CoA ,heterologous expression ,metabolic pathway ,Biotechnology ,TP248.13-248.65 - Abstract
Natural polyketides play important roles in clinical treatment, agriculture, and animal husbandry. Compared to natural hosts, heterologous chassis (especially Actinomycetes) have many advantages in production of polyketide compounds. As a widely studied model Actinomycete, Saccharopolyspora erythraea is an excellent host to produce valuable heterologous polyketide compounds. However, many host factors affect the expression efficiency of heterologous genes, and it is necessary to modify the host to adapt heterologous production. In this study, the CRISPR-Cas9 system was used to knock out the erythromycin biosynthesis gene cluster of Ab (erythromycin high producing stain). A fragment of 49491 bp in genome (from SACE_0715 to SACE_0733) was deleted, generating the recombinant strain AbΔery in which erythromycin synthesis was blocked and synthetic substrates methylmalonyl-CoA and propionyl-CoA accumulated enormously. Based on AbΔery as heterologous host, three genes, AsCHS, RgTAL, and Sc4CL, driven by strong promoters Pj23119, PermE, and PkasO, respectively, were introduced to produce novel polyketide by L-tyrosine and methylmalonyl-CoA. The product (E)-4-hydroxy-6-(4-hydroxystyryl)-3,5-dimethyl-2H-pyrone was identified in fermentation by LC-MS. High performance liquid chromatography analysis showed that knocking out ery BGC resulted in an increase of methylmalonyl-CoA by 142% and propionyl-CoA by 57.9% in AbΔery compared to WT, and the yield of heterologous product in AbΔery:AsCHS-RgTAL-Sc4CL was higher than WT:AsCHS-RgTAL-Sc4CL. In summary, this study showed that AbΔery could potentially serve as a precious heterologous host to boost the synthesis of other valuable polyketone compounds using methylmalonyl-CoA and propionyl-CoA in the future.
- Published
- 2021
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26. The Escherichia coli FadR transcription factor: Too much of a good thing?
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Cronan, John E.
- Subjects
- *
TRANSCRIPTION factors , *ESCHERICHIA coli , *ARCHAEBACTERIA , *INTRACELLULAR membranes , *CELL size , *CELL physiology - Abstract
Escherichia coli FadR is a transcription factor regulated by acyl‐CoA thioester binding that optimizes fatty acid (FA) metabolism in response to environmental FAs. FadR represses the fad genes of FA degradation (β‐oxidation) and activates the fab genes of FA synthesis thereby allowing E. coli to have its cake (acyl chains for phospholipid synthesis) and eat it (degrade acyl chains to acetyl‐CoA). Acyl‐CoA binding of FadR derepresses the transcription of the fad genes and cancels fab gene transcriptional activation. Activation of fab genes was thought restricted to the fabA and fabB genes of unsaturated FA synthesis, but FadR overproduction markedly increases yields of all FA acyl chains. Subsequently, almost all of the remaining fab genes were shown to be transcriptionally activated by FadR binding, but binding was very weak. Why are the low‐affinity sites retained? What effects on cell physiology would result from their conversion to high‐affinity sites (thereby mimicking FadR overproduction)? Investigations of E. coli cell size determinants showed that FA synthesis primarily determines E. coli cell size. Upon modest induction of FadR, cell size increases, but at the cost of growth rate and accumulation of intracellular membranes. Greater induction resulted in further growth rate decreases and abnormal cells. Hence, too much FadR is bad. FadR is extraordinarily conserved in γ‐proteobacteria but has migrated. Mycobacterium tuberculosis encodes FadR orthologs one of which is functional in E. coli. Strikingly, the FadR theme of acyl‐CoA‐dependent transcriptional regulation is found in a different transcription factor family where two Bacillus species plus bacterial and archaeal thermophiles contain related proteins of similar function. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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27. Isolation and characterization of Lipomyces starkeyi mutants with greatly increased lipid productivity following UV irradiation.
- Author
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Takaku, Hiroaki, Ebina, Sayaka, Kasuga, Kotoha, Sato, Rikako, Ara, Satoshi, Kazama, Haruka, Matsuzawa, Tomohiko, Yaoi, Katsuro, Araki, Hideo, Shida, Yosuke, Ogasawara, Wataru, Ishiya, Koji, Aburatani, Sachiyo, and Yamazaki, Harutake
- Subjects
- *
DENSITY gradient centrifugation , *PENTOSE phosphate pathway , *LIPIDS , *ULTRAVIOLET radiation , *ACYL coenzyme A , *VEGETABLE oils , *CENTRIFUGATION - Abstract
The oleaginous yeast Lipomyces starkeyi is an intriguing lipid producer that can produce triacylglycerol (TAG), a feedstock for biodiesel production. We previously reported that the L. starkeyi mutant E15 with high levels of TAG production compared with the wild-type was efficiently obtained using Percoll density gradient centrifugation. However, considering its use for biodiesel production, it is necessary to further improve the lipid productivity of the mutant. In this study, we aimed to obtain mutants with better lipid productivity than E15, evaluate its lipid productivity, and analyze lipid synthesis-related gene expression in the wild-type and mutant strains. The mutants E15-11, E15-15, and E15-25 exhibiting higher lipid productivity than E15 were efficiently isolated from cells exposed to ultraviolet light using Percoll density gradient centrifugation. They exhibited approximately 4.5-fold higher lipid productivity than the wild-type on day 3. The obtained mutants did not exhibit significantly different fatty acid profiles than the wild-type and E15 mutant strains. E15-11, E15-15, and E15-25 exhibited higher expression of acyl-CoA synthesis- and Kennedy pathway-related genes than the wild-type and E15 mutant strains. Activation of the pentose phosphate pathway, which supplies NADPH, was also observed. These results suggested that the increased expression of acyl-CoA synthesis- and Kennedy pathway-related genes plays a vital role in lipid productivity in the oleaginous yeast L. starkeyi. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
28. Acetyl-CoA synthetase (ACSS2) does not generate butyryl- and crotonyl-CoA.
- Author
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Zeaiter, Nour, Belot, Laura, Cunin, Valérie, Nahed, Roland Abi, Tokarska-Schlattner, Malgorzata, Le Gouellec, Audrey, Petosa, Carlo, Khochbin, Saadi, and Schlattner, Uwe
- Abstract
Acetyl and other acyl groups from different short-chain fatty acids (SCFA) competitively modify histones at various lysine sites. To fully understand the functional significance of such histone acylation, a key epigenetic mechanism, it is crucial to characterize the cellular sources of the corresponding acyl-CoA molecules required for the lysine modification. Like acetate, SCFAs such as propionate, butyrate and crotonate are thought to be the substrates used to generate the corresponding acyl-CoAs by enzymes known as acyl-CoA synthetases. The acetyl-CoA synthetase, ACSS2, which produces acetyl-CoA from acetate in the nucleocytoplasmic compartment, has been proposed to also mediate the synthesis of acyl-CoAs such as butyryl- and crotonyl-CoA from the corresponding SCFAs. This idea is now widely accepted and is sparking new research projects. However, based on our direct in vitro experiments with purified or recombinant enzymes and structural considerations, we demonstrate that ACSS2 is unable to mediate the generation of non-acetyl acyl-CoAs like butyryl- and crotonyl-CoA. It is therefore essential to re-examine published data and corresponding discussions in the light of this new finding. • No decreased histone butyrylation/crotonylation with ACSS2 knockdown. • ACSS2 has narrow substrate specificity. • Butyryl-/crotonyl-CoA are synthesized by enzymes different from ACSS2. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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- View/download PDF
29. Enhanced triacylglycerol production in the diatom Phaeodactylum tricornutum by inactivation of a Hotdog-fold thioesterase gene using TALEN-based targeted mutagenesis
- Author
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Xiahui Hao, Ling Luo, Juliette Jouhet, Fabrice Rébeillé, Eric Maréchal, Hanhua Hu, Yufang Pan, Xiaoming Tan, Zhuo Chen, Lingjie You, Hong Chen, Fang Wei, and Yangmin Gong
- Subjects
Acyl-CoA thioesterase ,Acyl-CoA ,Fatty acids ,Phaeodactylum tricornutum ,TALEN ,Triacylglycerols ,Fuel ,TP315-360 ,Biotechnology ,TP248.13-248.65 - Abstract
Abstract Background In photosynthetic oleaginous microalgae, acyl-CoA molecules are used as substrates for the biosynthesis of membrane glycerolipids, triacylglycerol (TAG) and other acylated molecules. Acyl-CoA can also be directed to beta-oxidative catabolism. They can be utilized by a number of lipid metabolic enzymes including endogenous thioesterases, which catalyze their hydrolysis to release free fatty acids. Acyl-CoA availability thus plays fundamental roles in determining the quantity and composition of membrane lipids and storage lipids. Results Here, we have engineered the model diatom Phaeodactylum tricornutum to produce significantly increased TAGs by disruption of the gene encoding a Hotdog-fold thioesterase involved in acyl-CoA hydrolysis (ptTES1). This plastidial thioesterase can hydrolyze both medium- and long-chain fatty acyl-CoAs, but has the highest activity toward long-chain saturated and monounsaturated fatty acyl-CoAs. The maximum rate was found with oleoyl-CoA, which is hydrolyzed at 50 nmol/min/mg protein. The stable and targeted interruption of acyl-CoA thioesterase gene was achieved using a genome editing technique, transcription activator-like effector nucleases (TALENs). Disruption of native ptTES1 gene resulted in a 1.7-fold increase in TAG content when algal strains were grown in nitrogen-replete media for 8 days, whereas the content of other lipid classes, including phosphoglycerolipids and galactoglycerolipids, remained almost unchanged. The engineered algal strain also exhibited a marked change in fatty acid profile, including a remarkable increase in 16:0 and 16:1 and a decrease in 20:5. Nitrogen deprivation for 72 h further increased TAG content and titer of the engineered strain, reaching 478 μg/109 cells and 4.8 mg/L, respectively. Quantitative determination of in vivo acyl-CoAs showed that the total acyl-CoA pool size was significantly higher in the engineered algal strain than that in the wild type. Conclusions This study supports the role of ptTES1 in free fatty acid homeostasis in the plastid of Phaeodactylum and demonstrates the potential of TALEN-based genome editing technique to generate an enhanced lipid-producing algal strain through blocking acyl-CoA catabolism.
- Published
- 2018
- Full Text
- View/download PDF
30. The Saccharomyces cerevisiae ABC subfamily D transporter Pxa1/Pxa2p co‐imports CoASH into the peroxisome.
- Author
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Roermund, Carlo W. T., IJlst, Lodewijk, Baker, Alison, Wanders, Ronald J. A., Theodoulou, Freddie L., and Waterham, Hans R.
- Subjects
- *
SACCHAROMYCES cerevisiae , *PEROXISOMES , *COENZYME A , *FATTY acids , *CELL metabolism - Abstract
ATP‐binding cassette (ABC) subfamily D transporters are important for the uptake of fatty acids and other beta‐oxidation substrates into peroxisomes. Genetic and biochemical evidence indicates that the transporters accept fatty acyl‐coenzyme A that is cleaved during the transport cycle and then re‐esterified in the peroxisomal lumen. However, it is not known whether free coenzyme A (CoA) is released inside or outside the peroxisome. Here we have used Saccharomyces cerevisiae and isolated peroxisomes to demonstrate that free CoA is released in the peroxisomal lumen. Thus, ABC subfamily D transporter provide an import pathway for free CoA that controls peroxisomal CoA homeostasis and tunes metabolism according to the cell's demands. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
31. Fatty acid metabolism in cyanobacteria
- Author
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Taylor, George and Smirnoff, Nicholas
- Subjects
500 ,acyl-CoA ,biofuel ,ß-oxidation ,Cyanobacteria ,fatty acid ,metabolism - Abstract
With crude oil demand rising and supplies being depleted, alternative energy, specifically biofuels, are of intense scientific interest. Current plant crop based biofuels suffer from several problems, most importantly the use of land needed for food. Cyanobacteria offer a solution to this problem as they do not compete with land for food and produce hydrocarbons that can be used as biofuels. Upon examination of metabolic pathways competing with hydrocarbon synthesis, it appeared that cyanobacteria lacked the major fatty acid degradative metabolic pathway β-oxidation, generally thought to be a universally occurring pathway. Lack of this pathway in cyanobacteria was confirmed by employing a range of analytical techniques. Bioinformatic analysis suggested that potential enzymes with β-oxidation activity were involved in other metabolic pathways. A sensitive assay was set up to detect acyl- CoAs, the substrates of β-oxidation, using liquid chromatography triple quadrupole mass spectrometry. None could be detected in cyanobacteria. No enzymatic activity from the rate-limiting acyl-CoA dehydrogenase/oxidase could be detected in cyanobacterial extracts. It was found that radiolabeled fatty acids fed to cyanobacteria were utilised for lipid membranes as opposed to being converted to CO2 by respiration or into other compounds by the TCA cycle. An element of the β-oxidation pathway, E. coli acyl-CoA synthetase was ectopically expressed in a strain of cyanobacteria and implications of the introduction of acyl-CoA synthesis were assessed. Finally, the regulation of the fatty acid biosynthetic pathway was investigated. It was determined that under conditions of excess fatty acid, the transcription of acetyl-CoA carboxylase and enoyl-ACP reductase was repressed and acyl-ACP synthetase involved in fatty acid recycling was induced. These results were discussed in relation to fatty acid oxidation and hydrocarbon biosynthesis in other organisms.
- Published
- 2012
32. Kinetic, Structural, and Mutational Analysis of Acyl-CoA Carboxylase From Thermobifida fusca YX
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Kiran-Kumar Shivaiah, Bryon Upton, and Basil J. Nikolau
- Subjects
acyl-CoA ,biotin-dependent carboxylases ,Thermobifida fusca YX ,site-directed mutagenesis ,enzyme kinetics ,Biology (General) ,QH301-705.5 - Abstract
Acyl-CoA carboxylases (AcCCase) are biotin-dependent enzymes that are capable of carboxylating more than one short chain acyl-CoA substrate. We have conducted structural and kinetic analyses of such an AcCCase from Thermobifida fusca YX, which exhibits promiscuity in carboxylating acetyl-CoA, propionyl-CoA, and butyryl-CoA. The enzyme consists of two catalytic subunits (TfAcCCA and TfAcCCB) and a non-catalytic subunit, TfAcCCE, and is organized in quaternary structure with a A6B6E6 stoichiometry. Moreover, this holoenzyme structure appears to be primarily assembled from two A3 and a B6E6 subcomplexes. The role of the TfAcCCE subunit is to facilitate the assembly of the holoenzyme complex, and thereby activate catalysis. Based on prior studies of an AcCCase from Streptomyces coelicolor, we explored whether a conserved Asp residue in the TfAcCCB subunit may have a role in determining the substrate selectivity of these types of enzymes. Mutating this D427 residue resulted in alterations in the substrate specificity of the TfAcCCase, increasing proficiency for carboxylating acetyl-CoA, while decreasing carboxylation proficiency with propionyl-CoA and butyryl-CoA. Collectively these results suggest that residue D427 of AcCCB subunits is an important, but not sole determinant of the substrate specificity of AcCCase enzymes.
- Published
- 2021
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- View/download PDF
33. Benzoyl-CoA conjugate accumulation as an initiating event for male reprotoxic effects in the rat? Structure–activity analysis, species specificity, and in vivo relevance.
- Author
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Laue, Heike, Badertscher, Remo P., Hostettler, Lu, Weiner-Sekiya, Yumiko, Haupt, Tina, Nordone, Adrian, Adamson, Gregory M., and Natsch, Andreas
- Subjects
- *
SPECIES specificity , *RATS , *BENZOIC acid , *STRUCTURE-activity relationships , *LIVER cells - Abstract
A number of para-substituted benzoic acids (p-BA) and chemicals metabolized to p-BA have been found to confer adverse effects in male rats on sperm viability, motility, and morphology. These effects are putatively associated with the metabolism of p-BA to toxic intermediates. We had shown that p-BA lead to accumulation of high levels of p-alkyl-benzoyl-CoA conjugates in plated primary rat hepatocytes. Here we further investigated the relevance of this metabolic pathway for the reprotoxic effects in rats and rabbits. We extended the structure–activity relationship to a set of 19 chemicals (nine reprotoxic and ten non-reprotoxic) and confirmed a very strong correlation between p-alkyl-benzoyl-CoA accumulation in rat hepatocytes and the toxic outcome. Species specificity was probed by comparing rat, rabbit and human hepatocytes, and p-benzoyl-CoA accumulation was found to be specific to the rat hepatocytes, not occurring in human hepatocytes. There was also very limited accumulation in hepatocytes from rabbits that are a non-responder species in in vivo studies. Tissues of rats treated with 3-(4-isopropylphenyl)-2-methylpropanal were analysed and p-isopropyl-benzoyl-CoA conjugates were detected in the liver and in the testes in animals at toxic doses indicating that the metabolism observed in vitro is relevant to the in vivo situation and the critical metabolite does also occur in the reproductive tissue. These multiple lines of evidence further support benzoyl-CoA accumulation as a key initiating event for a specific group of male reproductive toxicants, and indicate a species-specific effect in the rat. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
34. Compartmentalised acyl-CoA metabolism and roles in chromatin regulation
- Author
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Sophie Trefely, Claudia D. Lovell, Nathaniel W. Snyder, and Kathryn E. Wellen
- Subjects
Compartmentalisation ,Acyl-CoA ,Histone ,Acylation ,Metabolism ,Internal medicine ,RC31-1245 - Abstract
Background: Many metabolites serve as important signalling molecules to adjust cellular activities and functions based on nutrient availability. Links between acetyl-CoA metabolism, histone lysine acetylation, and gene expression have been documented and studied over the past decade. In recent years, several additional acyl modifications to histone lysine residues have been identified, which depend on acyl-coenzyme A thioesters (acyl-CoAs) as acyl donors. Acyl-CoAs are intermediates of multiple distinct metabolic pathways, and substantial evidence has emerged that histone acylation is metabolically sensitive. Nevertheless, the metabolic sources of acyl-CoAs used for chromatin modification in most cases remain poorly understood. Elucidating how these diverse chemical modifications are coupled to and regulated by cellular metabolism is important in deciphering their functional significance. Scope of review: In this article, we review the metabolic pathways that produce acyl-CoAs, as well as emerging evidence for functional roles of diverse acyl-CoAs in chromatin regulation. Because acetyl-CoA has been extensively reviewed elsewhere, we will focus on four other acyl-CoA metabolites integral to major metabolic pathways that are also known to modify histones: succinyl-CoA, propionyl-CoA, crotonoyl-CoA, and butyryl-CoA. We also briefly mention several other acyl-CoA species, which present opportunities for further research; malonyl-CoA, glutaryl-CoA, 3-hydroxybutyryl-CoA, 2-hydroxyisobutyryl-CoA, and lactyl-CoA. Each acyl-CoA species has distinct roles in metabolism, indicating the potential to report shifts in the metabolic status of the cell. For each metabolite, we consider the metabolic pathways in which it participates and the nutrient sources from which it is derived, the compartmentalisation of its metabolism, and the factors reported to influence its abundance and potential nuclear availability. We also highlight reported biological functions of these metabolically-linked acylation marks. Finally, we aim to illuminate key questions in acyl-CoA metabolism as they relate to the control of chromatin modification. Major conclusions: A majority of acyl-CoA species are annotated to mitochondrial metabolic processes. Since acyl-CoAs are not known to be directly transported across mitochondrial membranes, they must be synthesized outside of mitochondria and potentially within the nucleus to participate in chromatin regulation. Thus, subcellular metabolic compartmentalisation likely plays a key role in the regulation of histone acylation. Metabolite tracing in combination with targeting of relevant enzymes and transporters will help to map the metabolic pathways that connect acyl-CoA metabolism to chromatin modification. The specific function of each acyl-CoA may be determined in part by biochemical properties that affect its propensity for enzymatic versus non-enzymatic protein modification, as well as the various enzymes that can add, remove and bind each modification. Further, competitive and inhibitory effects of different acyl-CoA species on these enzymes make determining the relative abundance of acyl-CoA species in specific contexts important to understand the regulation of chromatin acylation. An improved and more nuanced understanding of metabolic regulation of chromatin and its roles in physiological and disease-related processes will emerge as these questions are answered.
- Published
- 2020
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35. Acyl-Coa Thioesterases: A Rheostat That Controls Activated Fatty Acids Modulates Dengue Virus Serotype 2 Replication
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Laura A. St Clair, Stephanie A. Mills, Elena Lian, Paul S. Soma, Aritra Nag, Caroline Montgomery, Gabriela Ramirez, Nunya Chotiwan, Rebekah C. Gullberg, and Rushika Perera
- Subjects
lipids ,dengue virus ,acyl-CoA ,acyl-CoA thioesterase ,fatty acids ,membranes ,Microbiology ,QR1-502 - Abstract
During infection with dengue viruses (DENVs), the lipid landscape within host cells is significantly altered to assemble membrane platforms that support viral replication and particle assembly. Fatty acyl-CoAs are key intermediates in the biosynthesis of complex lipids that form these membranes. They also function as key signaling lipids in the cell. Here, we carried out loss of function studies on acyl-CoA thioesterases (ACOTs), a family of enzymes that hydrolyze fatty acyl-CoAs to free fatty acids and coenzyme A, to understand their influence on the lifecycle of DENVs. The loss of function of the type I ACOTs 1 (cytoplasmic) and 2 (mitochondrial) together significantly increased DENV serotype 2 (DENV2) viral replication and infectious particle release. However, isolated knockdown of mitochondrial ACOT2 significantly decreased DENV2 protein translation, genome replication, and infectious virus release. Furthermore, loss of ACOT7 function, a mitochondrial type II ACOT, similarly suppressed DENV2. As ACOT1 and ACOT2 are splice variants, these data suggest that functional differences and substrate specificities due to the location (cytosol and mitochondria, respectively) of these proteins may account for the differences in DENV2 infection phenotype. Additionally, loss of mitochondrial ACOT2 and ACOT7 expression also altered the expression of several ACOTs located in multiple organelle compartments within the cell, highlighting a complex relationship between ACOTs in the DENV2 virus lifecycle.
- Published
- 2022
- Full Text
- View/download PDF
36. Fatty acid oxidation promotes reprogramming by enhancing oxidative phosphorylation and inhibiting protein kinase C
- Author
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Zhaoyu Lin, Fei Liu, Peiliang Shi, Anying Song, Zan Huang, Dayuan Zou, Qin Chen, Jianxin Li, and Xiang Gao
- Subjects
Cpt1 ,Palmitoylcarnitine ,Oxidative phosphorylation ,Acyl-CoA ,Reprogramming ,Fatty acid oxidation ,Medicine (General) ,R5-920 ,Biochemistry ,QD415-436 - Abstract
Abstract Background Changes in metabolic pathway preferences are key events in the reprogramming process of somatic cells to induced pluripotent stem cells (iPSCs). The optimization of metabolic conditions can enhance reprogramming; however, the detailed underlying mechanisms are largely unclear. By comparing the gene expression profiles of somatic cells, intermediate-phase cells, and iPSCs, we found that carnitine palmitoyltransferase (Cpt)1b, a rate-limiting enzyme in fatty acid oxidation, was significantly upregulated in the early stage of the reprogramming process. Methods Mouse embryonic fibroblasts isolated from transgenic mice carrying doxycycline (Dox)-inducible Yamanaka factor constructs were used for reprogramming. Various fatty acid oxidation-related metabolites were added during the reprogramming process. Colony counting and fluorescence-activated cell sorting (FACS) were used to calculate reprogramming efficiency. Fatty acid oxidation-related metabolites were measured by liquid chromatography–mass spectrometry. Seahorse was used to measure the level of oxidative phosphorylation. Results We found that overexpression of cpt1b enhanced reprogramming efficiency. Furthermore, palmitoylcarnitine or acetyl-CoA, the primary and final products of Cpt1-mediated fatty acid oxidation, also promoted reprogramming. In the early reprogramming process, fatty acid oxidation upregulated oxidative phosphorylation and downregulated protein kinase C activity. Inhibition of protein kinase C also promoted reprogramming. Conclusion We demonstrated that fatty acid oxidation promotes reprogramming by enhancing oxidative phosphorylation and inhibiting protein kinase C activity in the early stage of the reprogramming process. This study reveals that fatty acid oxidation is crucial for the reprogramming efficiency.
- Published
- 2018
- Full Text
- View/download PDF
37. Phosphorylation of the acyl-CoA binding pocket of the FadR transcription regulator in Sulfolobus acidocaldarius.
- Author
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Maklad, Hassan R., Gutierrez, Gustavo J., Esser, Dominik, Siebers, Bettina, and Peeters, Eveline
- Subjects
- *
ACYL coenzyme A , *PHOSPHORYLATION , *CHROMOSOME structure , *LIGAND binding (Biochemistry) , *TRANSCRIPTION factors , *ACYLTRANSFERASES - Abstract
The archaeal model organism Sulfolobus acidocaldarius possesses a TetR-like transcription factor that represses a 30-kb gene cluster encoding fatty acid metabolism enzymes. Interaction of this regulator, FadR Sa , with acyl-CoA molecules causes a DNA dissociation, which may lead to a derepression of the gene cluster. Previously, a phosphoproteome analysis revealed the phosphorylation of three consecutive amino acids in the acyl-CoA ligand binding pocket. Here, we study this phosphorylation event and show that ArnC, a Hanks-type protein kinase, targets a threonine within the phosphoacceptor motif in vitro. Electrophoretic mobility shift assays using a phosphomimetic mutant of FadR Sa demonstrate that the presence of negatively charged groups on the phosphoacceptor motif causes an inhibition of the ligand binding that desensitizes the responsiveness of the regulator to acyl-CoA molecules. Based on these observations, we propose a model in which phosphorylation of FadR Sa in its ligand-binding pocket acts as an additional regulatory layer silencing acyl-CoA responsive derepression of fatty acid and lipid degradation. Moreover, given the recently discovered interplay between FadR Sa and the chromosome structuring protein coalescin, FadR Sa phosphorylation could also influence local chromosome conformation under specific cellular conditions. • FadR is phosphorylated within its acyl-CoA-binding pocket. • The eukaryotic-type kinase ArnC phosphorylates FadR in vitro at threonine 134. • The phosphomimetic mutant of FadR is less sensitive for acyl-CoA. • FadR phosphorylation may control metabolism and chromatin conformation. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
38. Long-chain acyl-CoA synthetase 4 participates in the formation of highly unsaturated fatty acid-containing phospholipids in murine macrophages.
- Author
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Kuwata, Hiroshi, Nakatani, Eriko, Shimbara-Matsubayashi, Satoko, Ishikawa, Fumihiro, Shibanuma, Motoko, Sasaki, Yuka, Yoda, Emiko, Nakatani, Yoshihito, and Hara, Shuntaro
- Subjects
- *
ACYLTRANSFERASES , *PHOSPHOLIPIDS , *ACYL coenzyme A , *LIQUID chromatography-mass spectrometry , *UNSATURATED fatty acids , *MASS analysis (Spectrometry) , *FREE fatty acids - Abstract
Long-chain acyl-coenzyme A synthetases (ACSLs) are a family of enzymes that convert free long-chain fatty acids into their acyl-coenzyme A (CoA) forms. ACSL4, belonging to the ACSL family, shows a preferential use of arachidonic acid (AA) as its substrate and plays a role in the remodeling of AA-containing phospholipids by incorporating free AA. However, little is known about the roles of ACSL4 in inflammatory responses. Here, we assessed the roles of ACSL4 on the effector functions of bone marrow-derived macrophages (BMDMs) obtained from mice lacking ACSL4. Liquid chromatography–tandem mass spectrometry analysis revealed that various highly unsaturated fatty acid (HUFA)-derived fatty acyl-CoA species were markedly decreased in the BMDMs obtained from ACSL4-deficient mice compared with those in the BMDMs obtained from wild-type mice. BMDMs from ACSL4-deficient mice also showed a reduced incorporation of HUFA into phosphatidylcholines. The stimulation of BMDMs with lipopolysaccharide (LPS) elicited the release of prostaglandins (PGs), such as PGE 2 , PGD 2 and PGF 2α , and the production of these mediators was significantly enhanced by ACSL4 deficiency. In contrast, neither the LPS-induced release of cytokines, such as IL-6 and IL-10, nor the endocytosis of zymosan or dextran was affected by ACSL4 deficiency. These results suggest that ACSL4 has a crucial role in the maintenance of HUFA composition of certain phospholipid species and in the incorporation of free AA into the phospholipids in LPS-stimulated macrophages. ACSL4 dysfunction may facilitate inflammatory responses by an enhanced eicosanoid storm. • ACSL4 uses AA, EPA, adrenic acid, or DHA, as good substrates. • ACSL4-deficiency decreases the levels of PUFA-derived acyl-CoA in BMDMs. • High levels of PUFAs are observed in the culture medium of ACSL4 KO BMDMs. • ACSL4 enzyme competes with the COX enzymes for AA. • ACSL4 deficiency leads to substrate shunting towards PG production in BMDMs. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
39. Development of targeted hydrophilic interaction liquid chromatography-tandem mass spectrometry method for acyl-Coenzyme A covering short- to long-chain species in a single analytical run.
- Author
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Singh, Madhulika, Kiyuna, Ligia Akemi, Odendaal, Christoff, Bakker, Barbara M., Harms, Amy C., and Hankemeier, Thomas
- Subjects
- *
LIQUID chromatography-mass spectrometry , *HYDROPHILIC interaction liquid chromatography , *ACETYLCOENZYME A , *HYDROPHILIC interactions , *FATTY acid oxidation , *ACYL coenzyme A , *MATRIX effect , *MASS transfer coefficients - Abstract
• HILIC-MS/MS method covers free CoA and acyl-CoAs (short to long-chain) in one run. • The method employing a zwitterionic HILIC column was characterized. • Limit of detection is in the range of (1.3–12.4) pmol mL−1 in the targeted method. • Application in HepG2 cells shows high acyl-CoAs and low free CoA in starved state. Acyl-CoAs play a significant role in numerous physiological and metabolic processes making it important to assess their concentration levels for evaluating metabolic health. Considering the important role of acyl-CoAs, it is crucial to develop an analytical method that can analyze these compounds. Due to the structural variations of acyl-CoAs, multiple analytical methods are often required for comprehensive analysis of these compounds, which increases complexity and the analysis time. In this study, we have developed a method using a zwitterionic HILIC column that enables the coverage of free CoA and short- to long-chain acyl-CoA species in one analytical run. Initially, we developed the method using an LC-QTOF instrument for the identification of acyl-CoA species and optimizing their chromatography. Later, a targeted HILIC-MS/MS method was created in scheduled multiple reaction monitoring mode using a QTRAP MS detector. The performance of the method was evaluated based on various parameters such as linearity, precision, recovery and matrix effect. This method was applied to identify the difference in acyl-CoA profiles in HepG2 cells cultured in different conditions. Our findings revealed an increase in levels of acetyl-CoA, medium- and long-chain acyl-CoA while a decrease in the profiles of free CoA in the starved state, indicating a clear alteration in the fatty acid oxidation process. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
40. Mito-Nuclear Communication by Mitochondrial Metabolites and Its Regulation by B-Vitamins
- Author
-
Joëlle J. E. Janssen, Sander Grefte, Jaap Keijer, and Vincent C. J. de Boer
- Subjects
B-vitamins ,mito-nuclear signaling ,TCA cycle metabolites ,acyl-CoA ,reactive oxygen species ,sirtuins ,Physiology ,QP1-981 - Abstract
Mitochondria are cellular organelles that control metabolic homeostasis and ATP generation, but also play an important role in other processes, like cell death decisions and immune signaling. Mitochondria produce a diverse array of metabolites that act in the mitochondria itself, but also function as signaling molecules to other parts of the cell. Communication of mitochondria with the nucleus by metabolites that are produced by the mitochondria provides the cells with a dynamic regulatory system that is able to respond to changing metabolic conditions. Dysregulation of the interplay between mitochondrial metabolites and the nucleus has been shown to play a role in disease etiology, such as cancer and type II diabetes. Multiple recent studies emphasize the crucial role of nutritional cofactors in regulating these metabolic networks. Since B-vitamins directly regulate mitochondrial metabolism, understanding the role of B-vitamins in mito-nuclear communication is relevant for therapeutic applications and optimal dietary lifestyle. In this review, we will highlight emerging concepts in mito-nuclear communication and will describe the role of B-vitamins in mitochondrial metabolite-mediated nuclear signaling.
- Published
- 2019
- Full Text
- View/download PDF
41. Cholesterol-Mediated Coenzyme A Depletion in Catabolic Mutants of Mycobacteria Leads to Toxicity.
- Author
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Brown KL, Krekhno JMC, Xing S, Huan T, and Eltis LD
- Subjects
- Chromatography, Liquid, Cholesterol metabolism, Coenzyme A metabolism, Tandem Mass Spectrometry, Mycobacterium tuberculosis
- Abstract
Cholesterol is a critical growth substrate for Mycobacterium tuberculosis (Mtb) during infection, and the cholesterol catabolic pathway has been targeted for the development of new antimycobacterial agents. A key metabolite in cholesterol catabolism is 3aα-H-4α(3'-propanoate)-7aβ-methylhexahydro-1,5-indanedione (HIP). Many of the HIP metabolites are acyl-coenzyme A (CoA) thioesters, whose accumulation in deletion mutants can cause cholesterol-mediated toxicity. We used LC-MS/MS analysis to demonstrate that deletion of genes involved in HIP catabolism leads to acyl-CoA accumulation with concomitant depletion of free CoASH, leading to dysregulation of central metabolic pathways. CoASH and acyl-CoAs inhibited PanK, the enzyme that catalyzes the first step in the transformation of pantothenate to CoASH. Inhibition was competitive with respect to ATP with K values ranging from 9 μM for CoASH to 57 μM for small acyl-CoAs and 180 ± 30 μM for cholesterol-derived acyl-CoA. These findings link two critical metabolic pathways and suggest that therapeutics targeting cholesterol catabolic enzymes could both prevent the utilization of an important growth substrate and simultaneously sequester CoA from essential cellular processes, leading to bacterial toxicity.
ic values ranging from 9 μM for CoASH to 57 μM for small acyl-CoAs and 180 ± 30 μM for cholesterol-derived acyl-CoA. These findings link two critical metabolic pathways and suggest that therapeutics targeting cholesterol catabolic enzymes could both prevent the utilization of an important growth substrate and simultaneously sequester CoA from essential cellular processes, leading to bacterial toxicity.- Published
- 2024
- Full Text
- View/download PDF
42. Expression, purification, and characterization of a metagenomic thioesterase from activated sludge involved in the degradation of acylCoA-derivatives.
- Author
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Folch-Mallol, Jorge Luis, Zárate, Andrés, Sánchez-Reyes, Ayixón, and López- Lara, Isabel M.
- Subjects
- *
THIOESTERASE , *ACYLTRANSFERASES , *WATER treatment plant residuals , *XENOBIOTICS , *PHENYLACETIC acid - Abstract
Abstract Metagenomic libraries are a novel and powerful approach to seek for pathways involved in xenobiotic degradation, since this technique abolishes the need for cultivating microorganisms that otherwise would be overlooked if they cannot grow on standard laboratory media and conditions. In this paper, we describe the expression, purification and characterization of a novel metagenomic thioesterase which was described to be involved in phenylacetic acid degradation (A. Sánchez-Reyes, R. Batista-García, G. Valdés-García E. Ortiz, L. Perezgasga, A. Zárate-Romero, N. Pastor, J. L. Folch-Mallol, A Family 13 thioesterase isolated from an activated sludge metagenome: insights into aromatic compounds metabolism, Proteins 85 (2017) 1222–1237). According to similarity and phylogenetic analyses, the enzyme seems to belong to an Actinobacterium. Nevertheless, after a process of denaturation and refolding, the protein expressed in E. coli was obtained in an active form. New data concerning the substrate preferences for this enzyme are presented which suggest that this thioesterase could be involved in breaking the ester bond in the CoA-linear acyl derivatives of the phenylacetic acetic pathway. Highlights • Activated sludge from a water treatment plant as a source of activities to study xenobiotic compound degradation. • Characterization of substrate affinity for linear and Acyl-CoA derivatives by a metagenomic thioesterase purified from E.coli. • Actinobacterium protein expressed in E. coli and purified by solubilization of inclusion bodies and re-folding. • Metagenomic thioesterase involved in aromatic degradation pathways could be useful for bioremediation. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
43. Mito-Nuclear Communication by Mitochondrial Metabolites and Its Regulation by B-Vitamins.
- Author
-
Janssen, Joëlle J. E., Grefte, Sander, Keijer, Jaap, and de Boer, Vincent C. J.
- Subjects
MITOCHONDRIAL pathology ,HOMEOSTASIS ,ETIOLOGY of diseases ,METABOLITES ,VITAMIN B deficiency - Abstract
Mitochondria are cellular organelles that control metabolic homeostasis and ATP generation, but also play an important role in other processes, like cell death decisions and immune signaling. Mitochondria produce a diverse array of metabolites that act in the mitochondria itself, but also function as signaling molecules to other parts of the cell. Communication of mitochondria with the nucleus by metabolites that are produced by the mitochondria provides the cells with a dynamic regulatory system that is able to respond to changing metabolic conditions. Dysregulation of the interplay between mitochondrial metabolites and the nucleus has been shown to play a role in disease etiology, such as cancer and type II diabetes. Multiple recent studies emphasize the crucial role of nutritional cofactors in regulating these metabolic networks. Since B-vitamins directly regulate mitochondrial metabolism, understanding the role of B-vitamins in mito-nuclear communication is relevant for therapeutic applications and optimal dietary lifestyle. In this review, we will highlight emerging concepts in mito-nuclear communication and will describe the role of B-vitamins in mitochondrial metabolite-mediated nuclear signaling. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
44. Low-oxygen response is triggered by an ATP-dependent shift in oleoyl-CoA in Arabidopsis.
- Author
-
Schmidt, Romy R., Fulda, Martin, Paul, Melanie V., Anders, Max, Plum, Frederic, Weits, Daniel A., Kosmacz, Monika, Larsone, Tony R., Graham, Ian A., Beemster, Gerrit T. S., Licausi, Francesco, Geigenberger, Peter, Schippers, Jos H., and van Dongena, Joost T.
- Subjects
- *
REACTIVE oxygen species , *GENE expression , *MOLECULAR biology , *ARABIDOPSIS , *TRANSCRIPTION factors - Abstract
Plant response to environmental stimuli involves integration of multiple signals. Upon low-oxygen stress, plants initiate a set of adaptive responses to circumvent an energy crisis. Here, we reveal how these stress responses are induced by combining (i) energydependent changes in the composition of the acyl-CoA pool and (ii) the cellular oxygen concentration. A hypoxia-induced decline of cellular ATP levels reduces LONG-CHAIN ACYL-COA SYNTHETASE activity, which leads to a shift in the composition of the acyl-CoA pool. Subsequently, we show that different acyl-CoAs induce unique molecular responses. Altogether, our data disclose a role for acyl-CoAs acting in a cellular signaling pathway in plants. Upon hypoxia, high oleoyl-CoA levels provide the initial trigger to release the transcription factor RAP2.12 from its interaction partner ACYL-COA BINDING PROTEIN at the plasma membrane. Subsequently, according to the N-end rule for proteasomal degradation, oxygen concentrationdependent stabilization of the subgroup VII ETHYLENE-RESPONSE FACTOR transcription factor RAP2.12 determines the level of hypoxia-specific gene expression. This research unveils a specific mechanism activating low-oxygen stress responses only when a decrease in the oxygen concentration coincides with a drop in energy. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
45. The Role of Acyl-CoA Pools and Metabolic Adaptations in Response to Altered Lipid Metabolism
- Author
-
Pearce, Ryan William
- Subjects
- Biochemistry, Analytical Chemistry, Chemistry, Medicine, fabry disease, lysosomal storage disorder, acyl-coa, coenzyme a, lc-ms, mass spectrometry, metabolomics, qtof, fibroblast model
- Abstract
The integration of acyl-CoAs into multiple pathways demonstrates their utility as biomarkers for the study of fatty acid metabolism, particularly in the context of physiological states which affect energy production. Fabry disease is an inherited lysosomal storage disorder that affects multiple organ systems including the heart, for which the underlying mechanisms are poorly understood. A portfolio of LC-MS/MS based targeted and high-resolution untargeted assays were developed to measure key metabolites including acyl-CoAs, acylcarnitines, and tricarboxylic acid cycle intermediates. These methods were used to study a cellular model of Fabry disease using fibroblasts from donors representing 2 different phenotypes (GM00107 & GM00882, Coriell Institute). Stable isotope tracer analysis of 13C2-acetyl-CoA enrichment from fully 13C labeled glucose and palmitate revealed markedly reduced 13C2-acetyl-CoA labeling from both palmitate and glucose in GM00107 Fabry cell line compared to healthy controls, suggesting that there is severe mitochondrial dysfunction preventing normal oxidative metabolism of primary carbon substrates. Untargeted metabolomics revealed a significant decrease of ATP and phosphocreatine in Fabry disease. Therefore, the accumulation of glycosphingolipids in lysosomes may disrupt the cellular homeostasis and energy production of the cell contributing to mitochondrial dysfunction and a shift towards compensatory mechanisms in Fabry disease.
- Published
- 2023
46. Recent developments in the analytical approaches of acyl-CoAs to assess their role in mitochondrial fatty acid oxidation disorders.
- Author
-
Singh, Madhulika, Elfrink, Hyung L., Harms, Amy C., and Hankemeier, Thomas
- Subjects
- *
FATTY acid oxidation , *INBORN errors of metabolism , *LIQUID chromatography-mass spectrometry , *CARRIER proteins , *ENZYME deficiency , *ACYLTRANSFERASES - Abstract
Fatty acid oxidation disorders (FAOD) are inborn errors of metabolism that occur due to deficiency of specific enzyme activities and transporter proteins involved in the mitochondrial metabolism of fatty acids, causing a deficiency in ATP production. The identification of suitable biomarkers plays a crucial role in predicting the future risk of disease and monitoring responses to therapies. Acyl-CoAs are directly involved in the steps of fatty acid oxidation and are the primary biomarkers associated with FAOD. However, acyl-CoAs are not used as diagnostic biomarkers in hospitals and clinics as they are present intracellularly with low endogenous levels. Additionally, the analytical method development of acyl-CoAs is quite challenging due to diverse physicochemical properties and instability. Hence, secondary biomarkers such as acylcarnitines are used for the identification of FAOD. In this review, the focus is on the analytical techniques that have evolved over the years for the identification and quantitation of acyl-CoAs. Among these techniques, liquid chromatography-mass spectrometry clearly has an advantage in terms of sensitivity and selectivity. Stable isotope labeling by essential nutrients in cell culture (SILEC) enables the generation of labeled internal standards. Each acyl-CoA species has a distinct pattern of instability and degradation, and the use of appropriately matched internal standards can compensate for such issues. Although significant progress has been made in measuring acyl-CoAs, more efforts are needed for bringing these technical advancements to hospitals and clinics. This review also highlights the difficulties involved in the routine use of acyl-CoAs as a diagnostic biomarker and some of the measures that can be adopted by clinics and hospitals for overcoming these limitations. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
47. Effect of Single Nucleotide Polymorphism rs1044925 in Acyl-CoA: Cholesterol Acyltransferase-1 Gene on Plasma Lipid Parameters in Patients with Ischemic Stroke
- Author
-
Johnson Oshiobugie Momoh
- Subjects
medicine.medical_specialty ,Geography, Planning and Development ,Sterol O-acyltransferase ,Single-nucleotide polymorphism ,Development ,Acyl-CoA ,chemistry.chemical_compound ,Endocrinology ,chemistry ,Internal medicine ,Plasma lipids ,Ischemic stroke ,medicine ,SNP ,lipids (amino acids, peptides, and proteins) ,In patient ,Gene - Abstract
Acyl-coenzyme A: cholesterol acyltransferase-1 (ACAT-1) catalyzes the synthesis of cholesteryl esters from cholesterol and fatty acyl-CoA in tissues and the enzyme plays a major role in atherosclerosis and cellular cholesterol homeostasis. The study shows the effect of single nucleotide polymorphism rs1044925 in acyl-CoA:cholesterol acyltransferase-1 gene on plasma lipid parameters in patients with ischemic stroke. 100 patients with ischemic stroke and 100 controls matched for sex and aged 46-87 were selected for the study. Lipid profiles were measured using Randox kits and lipoprotein ratios were calculated using Excel software. The genotyping of the acyl-coenzyme A: cholesterol acyltransferase-1 rs1044925 SNP were performed by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) combined with 2% gel electrophoresis. There were significant difference (P
- Published
- 2021
48. H2O2 release from the very long chain acyl-CoA dehydrogenase
- Author
-
Pâmela A.H.B. Kakimoto, Fábio K. Tamaki, Ariel R. Cardoso, Sandro R. Marana, and Alicia J. Kowaltowski
- Subjects
Mitochondria ,Hydrogen peroxide ,Fatty acid oxidation ,Acyl-CoA ,Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
Enhanced mitochondrial generation of oxidants, including hydrogen peroxide (H2O2), is related to a large number of pathological conditions, including diet-induced obesity and steatohepatosis. Indeed, we have previously shown that high fat diets increase the generation of H2O2 in liver mitochondria energized by activated fatty acids. Here, we further study fatty-acid induced H2O2 release in liver mitochondria, and determine the characteristics that regulate it. We find that this production of H2O2 is independent of mitochondrial inner membrane integrity and insensitive to purine nucleotides. On the other hand, palmitate-induced H2O2 production is strongly enhanced by high fat diets and is pH-sensitive, with a peak at a matrix pH of ~8.5. Using recombinantly expressed human very long chain acyl-CoA dehydrogenase, we are able to demonstrate that palmitate-induced H2O2 release may be ascribed to the activity of this enzyme alone, acting as an oxidase. Our results add to a number of findings indicating that sources outside of the electron transport chain can generate significant, physiopathologically relevant, amounts of oxidants in mitochondria.
- Published
- 2015
- Full Text
- View/download PDF
49. Glycerophosphate/Acylglycerophosphate Acyltransferases
- Author
-
Atsushi Yamashita, Yasuhiro Hayashi, Naoki Matsumoto, Yoko Nemoto-Sasaki, Saori Oka, Takashi Tanikawa, and Takayuki Sugiura
- Subjects
acyltransferase ,acyl-CoA ,triacylglycerol ,phospholipid ,GPAT ,AGPAT ,Biology (General) ,QH301-705.5 - Abstract
Acyl-CoA:glycerol-3-phosphate acyltransferase (GPAT) and acyl-CoA: 1-acyl-glycerol-3-phosphate acyltransferase (AGPAT) are involved in the de novo synthesis of triacylglycerol (TAG) and glycerophospholipids. Many enzymes belonging to the GPAT/AGPAT family have recently been identified and their physiological or pathophysiological roles have been proposed. The roles of GPAT/AGPAT in the synthesis of TAG and obesity-related diseases were revealed through the identification of causative genes of these diseases or analyses of genetically manipulated animals. Recent studies have suggested that some isoforms of GPAT/AGPAT family enzymes are involved in the fatty acid remodeling of phospholipids. The enzymology of GPAT/AGPAT and their physiological/ pathological roles in the metabolism of glycerolipids have been described and discussed in this review.
- Published
- 2014
- Full Text
- View/download PDF
50. Comprehensive quantitative analysis of fatty-acyl-Coenzyme A species in biological samples by ultra-high performance liquid chromatography–tandem mass spectrometry harmonizing hydrophilic interaction and reversed phase chromatography.
- Author
-
Abrankó, László, Williamson, Gary, Gardner, Samantha, and Kerimi, Asimina
- Subjects
- *
FATTY-acyl-CoA , *BIOMARKERS , *CELL metabolism , *HIGH performance liquid chromatography , *TANDEM mass spectrometry - Abstract
Fatty acyl-Coenzyme A species (acyl-CoAs) are key biomarkers in studies focusing on cellular energy metabolism. Existing analytical approaches are unable to simultaneously detect the full range of short-, medium-, and long-chain acyl-CoAs, while chromatographic limitations encountered in the analysis of limited amounts of biological samples are an often overlooked problem. We report the systematic development of a UHPLC–ESI-MS/MS method which incorporates reversed phase (RP) and hydrophilic interaction liquid chromatography (HILIC) separations in series, in an automated mode. The protocol outlined encompasses quantification of acyl-CoAs of varying hydrophobicity from C2 to C20 with recoveries in the range of 90–111 % and limit of detection (LOD) 1–5 fmol, which is substantially lower than previously published methods. We demonstrate that the poor chromatographic performance and signal losses in MS detection, typically observed for phosphorylated organic molecules, can be avoided by the incorporation of a 0.1% phosphoric acid wash step between injections. The methodological approach presented here permits a highly reliable, sensitive and precise analysis of small amounts of tissues and cell samples as demonstrated in mouse liver, human hepatic (HepG2) and skeletal muscle (LHCNM2) cells. The considerable improvements discussed pave the way for acyl-CoAs to be incorporated in routine targeted lipid biomarker profile studies. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
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