Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.997
Filtrar
1.
Food Chem ; 400: 134061, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36084591

RESUMO

The underlying mechanism of the role of mitochondria in color changing of tilapia fillet during 0-4 d storage is not completely clear. A total of 209 differentially significant expressed proteins (DSEPs) were identified by using label-free mitochondrial proteomics, with 56 proteins up-regulated in T2 and 61 proteins (up-regulated) in T3. Protein-Protein interaction reveled proteins which participate in TCA cycles (Citrate synthase (cs)), Oxidoreductase (Malate dehydrogenase (mdh1, mdh2), Succinyl-CoA (Oxct1), Hydroxyacyl-coenzyme a dehydrogenase (hadh), Dehydrogenase/reductase (SDR family) member 1 (dhrs1)) interacted strongly with each other. In turn, they can increase the level of mitochondrial respiration and mitochondrial function, leading to color changing of tilapia fillet. The heat shock 60kD protein 1 (chaperonin, hspd1) interacted with metabolic enzymes (cs and mdh2) and had important effects on color. These results could help researchers better understand the color changing mechanism on the surface of tilapia fillet during the storage.


Assuntos
Carne Vermelha , Tilápia , Animais , Citrato (si)-Sintase/metabolismo , Coenzima A , Malato Desidrogenase/genética , Malato Desidrogenase/metabolismo , Proteínas Mitocondriais , Proteômica , Carne Vermelha/análise , Tilápia/genética , Tilápia/metabolismo
2.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 34(4): 352-360, 2022 Aug 23.
Artigo em Chinês | MEDLINE | ID: mdl-36116924

RESUMO

OBJECTIVE: To screen differentially expressed genes (DEGs) associated with chronic schistosomiasis japonica-induced hepatic fibrosis and analyze their functions. METHODS: The dataset of gene expression profiles of patients with chronic schistosomiasis japonica-induced hepatic fibrosis was downloaded from the Gene Expression Omnibus (GEO) database, and DEGs were screened using R package. The biological functions of DEGs were characterized using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. In addition, the protein-protein interaction (PPI) network of DEGs was created to screen the hub genes. RESULTS: A total of 62 DEGs were identified, including 12 down-regulated genes and 50 up-regulated genes. GO enrichment analysis showed that DEGs were mainly enriched in 116 biological processes, including fatty acid, sulfur compound, acyl-coenzyme A and thioester metabolism; 19 cellular components, including mitochondrial matrix, outer mitochondrial membrane and organelle outer membrane; and 7 molecular functions, including insulin-like growth factor binding and oxidoreductase activity. KEGG pathway enrichment analysis that the DEGs were significantly enriched in phosphatidylinositol-3-kinase/serine/threonine protein kinase (PI3K/Akt), mitogen-activated protein kinase (MAPK), calcium metabolism and cyclic adenosine monophosphate (cAMP) signaling. PPI network analysis identified six hub genes involved in the development of chronic schistosomiasis japonica-induced hepatic fibrosis, including ACACA, ACSL1, GPAM, THRSP, PLIN1 and DGAT2, and ACSL1, ACACA and PLIN1 were the top 3 hub genes. CONCLUSIONS: ACSL1, ACACA and PLIN1 may be the hub genes associated with the development of chronic schistosomiasis japonica-induced hepatic fibrosis, and abnormal lipid metabolism mediated by these DEGs may play an important role in the development of chronic schistosomiasis japonica-induced hepatic fibrosis.


Assuntos
Esquistossomose Japônica , Somatomedinas , Monofosfato de Adenosina , Cálcio , Coenzima A , Biologia Computacional , Ácidos Graxos , Humanos , Cirrose Hepática/genética , Proteínas Quinases Ativadas por Mitógeno , Oxirredutases , Fosfatidilinositol 3-Quinases , Fosfatidilinositóis , Proteínas Proto-Oncogênicas c-akt , Esquistossomose Japônica/complicações , Esquistossomose Japônica/genética , Serina , Compostos de Enxofre , Treonina
3.
Drug Des Devel Ther ; 16: 3071-3085, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36118165

RESUMO

Background: Dihydromyricetin (DHM) exerts protective effects in various brain diseases. The aim of this research was to investigate the biological role of DHM in cerebral ischemia reperfusion (I/R) injury. Methods: We generated a rat model of cerebral I/R injury by performing middle cerebral artery occlusion/reperfusion (MCAO/R). The neurological score and brain water content of the experimental rats was then evaluated. The infarct volume and extent of apoptosis in brain tissues was then assessed by 2,3,5-triphenyltetrazolium (TTC) and TdT-mediated dUTP nick end labeling (TUNEL) staining. Hippocampal neuronal cells (HT22) were subjected to oxygen-glucose deprivation/reperfusion (OGD/R) and cell counting kit-8 (CCK-8) assays and flow cytometry were performed to detect cell viability and apoptosis. The levels of lipid reactive oxygen species (ROS) and iron were detected and the expression levels of key proteins were assessed by Western blotting. Results: DHM obviously reduced neurological deficits, brain water content, infarct volume and cell apoptosis in the brain tissues of MCAO/R rats. DHM repressed ferroptosis and inhibited the sphingosine kinase 1 (SPHK1)/mammalian target of rapamycin (mTOR) pathway in MCAO/R rats. In addition, DHM promoted cell viability and repressed apoptosis in OGD/R-treated HT22 cells. DHM also suppressed the levels of lipid ROS and intracellular iron in OGD/R-treated HT22 cells. The expression levels of glutathione peroxidase 4 (GPX4) was enhanced while the levels of acyl-CoA synthetase long-chain family member 4 (ACSL4) and phosphatidylethanolamine binding protein 1 (PEBP1) were reduced in OGD/R-treated HT22 cells in the presence of DHM. Moreover, the influence conferred by DHM was abrogated by the overexpression of SPHK1 or treatment with MHY1485 (an activator of mTOR). Conclusion: This research demonstrated that DHM repressed ferroptosis by inhibiting the SPHK1/mTOR signaling pathway, thereby alleviating cerebral I/R injury. Our findings suggest that DHM may be a candidate drug for cerebral I/R injury treatment.


Assuntos
Ferroptose , Traumatismo por Reperfusão , Animais , Coenzima A/metabolismo , Coenzima A/farmacologia , Coenzima A/uso terapêutico , Flavonóis , Glucose/farmacologia , Infarto da Artéria Cerebral Média/tratamento farmacológico , Infarto da Artéria Cerebral Média/metabolismo , Ferro , Ligases/metabolismo , Ligases/farmacologia , Ligases/uso terapêutico , Lipídeos/farmacologia , Mamíferos/metabolismo , Oxigênio/metabolismo , Proteína de Ligação a Fosfatidiletanolamina/metabolismo , Proteína de Ligação a Fosfatidiletanolamina/farmacologia , Proteína de Ligação a Fosfatidiletanolamina/uso terapêutico , Fosfolipídeo Hidroperóxido Glutationa Peroxidase , Fosfotransferases (Aceptor do Grupo Álcool) , Ratos , Espécies Reativas de Oxigênio/metabolismo , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/metabolismo , Transdução de Sinais , Sincalida/metabolismo , Sincalida/farmacologia , Sincalida/uso terapêutico , Serina-Treonina Quinases TOR/metabolismo , Água
4.
Proc Natl Acad Sci U S A ; 119(40): e2207505119, 2022 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-36161908

RESUMO

Mycobacterium abscessus, an opportunistic pathogen responsible for pulmonary infections, contains genes predicted to encode two steroid catabolic pathways: a cholesterol catabolic pathway similar to that of Mycobacterium tuberculosis and a 4-androstenedione (4-AD) catabolic pathway. Consistent with this prediction, M. abscessus grew on both steroids. In contrast to M. tuberculosis, Rhodococcus jostii RHA1, and other Actinobacteria, the cholesterol and 4-AD catabolic gene clusters of the M. abscessus complex lack genes encoding HsaD, the meta-cleavage product (MCP) hydrolase. However, M. abscessus ATCC 19977 harbors two hsaD homologs elsewhere in its genome. Only one of the encoded enzymes detectably transformed steroid metabolites. Among tested substrates, HsaDMab and HsaDMtb of M. tuberculosis had highest substrate specificities for MCPs with partially degraded side chains thioesterified with coenzyme A (kcat/KM = 1.9 × 104 and 5.7 × 103 mM-1s-1, respectively). Consistent with a dual role in cholesterol and 4-AD catabolism, HsaDMab also transformed nonthioesterified substrates efficiently, and a ΔhsaD mutant of M. abscessus grew on neither steroid. Interestingly, both steroids prevented growth of the mutant on acetate. The ΔhsaD mutant of M. abscessus excreted cholesterol metabolites with a fully degraded side chain, while the corresponding RHA1 mutant excreted metabolites with partially degraded side chains. Finally, the ΔhsaD mutant was not viable in macrophages. Overall, our data establish that the cholesterol and 4-AD catabolic pathways of M. abscessus are unique in that they converge upstream of where this occurs in characterized steroid-catabolizing bacteria. The data further indicate that cholesterol is a substrate for intracellular bacteria and that cholesterol-dependent toxicity is not strictly dependent on coenzyme A sequestration.


Assuntos
Mycobacterium abscessus , Mycobacterium tuberculosis , Tuberculose , Androstenodiona/metabolismo , Colesterol/metabolismo , Coenzima A/metabolismo , Humanos , Hidrolases/metabolismo , Mycobacterium abscessus/genética , Mycobacterium abscessus/metabolismo , Mycobacterium tuberculosis/metabolismo , Esteroides/metabolismo
5.
Hear Res ; 424: 108604, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36116178

RESUMO

Age-related hearing loss (ARHL) is a common sensory deficit in the elderly, which seriously affects physical and mental health. Therefore, understanding its underlying molecular mechanisms and taking interventions to treat ARHL are urgently needed. In our study, cochlea of 4-week-old C57BL/6 mice as the Youth group (n = 6) and 48-week-old cochlea as the Old group (n = 6) were subjected to quasi-targeted metabolomics analysis by Ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). In total, 208 differential metabolites were identified in 12 cochlea samples, which highlighted the following discriminant compounds: tryptophan, piperidine, methionine, L-arginine, histamine, serotonin, acetylcholine, and 4-aminobutyric acid. Differentially expressed metabolites were identified which were involved in KEGG pathways related to the digestion and absorption of oxidative stress associated amino acids, Synaptic vesicle cycle of serotonin, Pantothenate and CoA Biosynthesis. These findings are a first step toward elucidating the pathophysiological pathways involved in the etiology of ARHL and provide the possibility to further explore the mechanisms of ARHL using metabolomic analysis.


Assuntos
Presbiacusia , Espectrometria de Massas em Tandem , Acetilcolina , Aminoácidos , Animais , Arginina , Cromatografia Líquida de Alta Pressão/métodos , Coenzima A , Histamina , Metionina , Camundongos , Camundongos Endogâmicos C57BL , Piperidinas , Presbiacusia/diagnóstico , Serotonina , Espectrometria de Massas em Tandem/métodos , Triptofano , Ácido gama-Aminobutírico
6.
Hum Exp Toxicol ; 41: 9603271221125934, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36171180

RESUMO

The resistance of glioblastoma to chemotherapy remains a significant clinical problem. Targeting alternative pathways such as protein prenylation is known to be effective against many cancers. Fluvastatin is a potent competitive inhibitor of 3-hydroxy-3-methylglutaryl- CoA (HMG-CoA) reductase, thereby inhibits prenylation. We demonstrate that fluvastatin alone effectively inhibits proliferation and induces apoptosis in multiple human glioblastoma cell lines. The combination index analysis shows that fluvastatin acts synergistically with common chemotherapy drugs for glioblastoma: temozolomide and irinotecan. We further show that fluvastatin acts on glioblastoma through inhibiting prenylation-dependent Ras activation. The combination of fluvastatin and low dose temozolomide resulted in remarkable inhibition of glioblastoma tumor in mice throughout the whole treatment duration without causing toxicity. Such combinatorial effects provide the basis for utilizing these FDA-approved drugs as a potential clinical approach in overcoming resistance and improving glioblastoma treatment.


Assuntos
Glioblastoma , Inibidores de Hidroximetilglutaril-CoA Redutases , Animais , Coenzima A/farmacologia , Coenzima A/uso terapêutico , Resistencia a Medicamentos Antineoplásicos , Ácidos Graxos Monoinsaturados/farmacologia , Ácidos Graxos Monoinsaturados/uso terapêutico , Fluvastatina/farmacologia , Fluvastatina/uso terapêutico , Glioblastoma/tratamento farmacológico , Glioblastoma/patologia , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Irinotecano/farmacologia , Irinotecano/uso terapêutico , Camundongos , Oxirredutases , Prenilação de Proteína , Temozolomida/farmacologia
7.
Methods Mol Biol ; 2546: 83-94, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36127580

RESUMO

Acylcarnitines are formed when an acyl group is transferred from coenzyme A to a molecule of L-carnitine. In organic acidemias, and in fatty acid oxidation disorders, specific acylcarnitine species accumulate in a pattern that is characteristic for each disease. For this reason, acylcarnitine analysis is widely used for screening and diagnosis of inherited disorders of metabolism. The most common method for acylcarnitine analysis uses flow injection tandem mass spectrometry. Flow injection analysis allows for high throughput, however, does not provide separation of isomeric and isobaric compounds. Among the acylcarnitine species which can be affected by the presence of isomeric/isobaric compounds, C4-carnitine and C5DC-carnitine are probably the ones encountered most often. The method presented here is performed on urine and utilizes butanolic HCL to derivatize acylcarnitines, ultra-performance liquid chromatography to resolve C4- and C5-DC isomers and isobars, and quantitation of these species using multiple-reaction monitoring (MRM).


Assuntos
Carnitina , Espectrometria de Massas em Tandem , Carnitina/análogos & derivados , Carnitina/análise , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida/métodos , Coenzima A , Ácidos Graxos , Espectrometria de Massas em Tandem/métodos
8.
Methods Mol Biol ; 2546: 295-309, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36127599

RESUMO

Measurement of methylmalonic acid (MMA) plays an important role in the diagnosis of vitamin B12 deficiency. Vitamin B12 is an essential cofactor for the enzymatic carbon rearrangement of methylmalonyl-CoA (MMA-CoA) to succinyl-CoA (SA-CoA), and the lack of vitamin B12 leads to elevated concentrations of MMA. Measurement of MMA in biological samples is complicated because of the presence of succinic acid (SA), isomer of MMA. We developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for MMA. The method utilizes derivatization and positive ion mode ionization, which is specific to polycarboxylic acids (MMA and SA are dicarboxylic acids), while derivatives of monocarboxylic acids at these conditions are not ionizable and not detectable. The only organic acid, other than MMA, that is detected in this method is SA. The described method does not require chromatographic resolution of the peaks of MMA and SA; quantitative measurement of MMA is performed using a deconvolution algorithm, which mathematically resolves signal corresponding to MMA, from the combined signal of MMA/SA. Because of the high selectivity of detection, this method utilizes isocratic chromatographic separation; reconditioning and re-equilibration of the chromatographic column between injections is unnecessary. The above features allow high-throughput analysis of MMA with injection-to-injection cycle time of approximately 1 minute.


Assuntos
Ácido Metilmalônico , Espectrometria de Massas em Tandem , Carbono , Cromatografia Líquida/métodos , Coenzima A , Ácido Metilmalônico/química , Succinatos , Espectrometria de Massas em Tandem/métodos , Vitaminas
9.
Methods Mol Biol ; 2546: 509-521, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36127618

RESUMO

Peroxisomal disorders are a heterogeneous group of genetic disorders caused by impaired peroxisomal biogenesis or by defects in single peroxisomal proteins. The most common peroxisomal disorders are Zellweger spectrum disorders (ZSDs), due to pathogenic variants in one of the 13 PEX genes, and X-linked adrenoleukodystrophy/adrenomyeloneuropathy (X-ALD/AMN), due to pathogenic variants in ATP-binding cassette transporter type D1 (ABCD1) gene. Peroxisomes perform multiple essential cellular functions, including ß-oxidation of very-long-chain fatty acids (VLCFAs), pristanic acid and some bile acid intermediates, and α-oxidation of phytanic acid. In most patients, abnormal levels of VLCFAs and/or branched-chain fatty acids (BCFAs, e.g., phytanic and pristanic acids) are present; hence, measuring these analytes is critical when suspecting a peroxisomal disorder. This chapter describes a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify VLCFAs and BCFAs in plasma or serum for the diagnosis of peroxisomal disorders. The method consists of an acid hydrolysis step to release the fatty acids from their coenzyme A esters followed by derivatization using oxalyl chloride, dimethylaminoethanol, and then methyl iodide. The trimethyl-amino-ethyl (TMAE) iodide ester derivatives are analyzed using UPLC-MS/MS in positive electrospray ionization and multiple reaction-monitoring (MRM) mode. Quantitation is performed using a five-point calibration curve after normalizing with deuterated internal standards.


Assuntos
Adrenoleucodistrofia , Transtornos Peroxissômicos , Transportadores de Cassetes de Ligação de ATP/metabolismo , Adrenoleucodistrofia/diagnóstico , Adrenoleucodistrofia/metabolismo , Ácidos e Sais Biliares , Cromatografia Líquida , Coenzima A/metabolismo , Deanol , Ésteres , Ácidos Graxos/metabolismo , Humanos , Iodetos/metabolismo , Transtornos Peroxissômicos/diagnóstico , Transtornos Peroxissômicos/metabolismo , Ácido Fitânico , Espectrometria de Massas em Tandem/métodos
10.
Redox Biol ; 56: 102446, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36057161

RESUMO

AIMS: Metabolic switching during heart development contributes to postnatal cardiomyocyte (CM) cell cycle exit and loss of regenerative capacity in the mammalian heart. Metabolic control has potential for developing effective CM proliferation strategies. We sought to determine whether lactate dehydrogenase A (LDHA) regulated CM proliferation by inducing metabolic reprogramming. METHODS AND RESULTS: LDHA expression was high in P1 hearts and significantly decreased during postnatal heart development. CM-specific LDHA knockout mice were generated using CRISPR/Cas9 technology. CM-specific LDHA knockout inhibited CM proliferation, leading to worse cardiac function and a lower survival rate in the neonatal apical resection model. In contrast, CM-specific overexpression of LDHA promoted CM proliferation and cardiac repair post-MI. The α-MHC-H2B-mCh/CAG-eGFP-anillin system was used to confirm the proliferative effect triggered by LDHA on P7 CMs and adult hearts. Metabolomics, proteomics and Co-IP experiments indicated that LDHA-mediated succinyl coenzyme A reduction inhibited succinylation-dependent ubiquitination of thioredoxin reductase 1 (Txnrd1), which alleviated ROS and thereby promoted CM proliferation. In addition, flow cytometry and western blotting showed that LDHA-driven lactate production created a beneficial cardiac regenerative microenvironment by inducing M2 macrophage polarization. CONCLUSIONS: LDHA-mediated metabolic reprogramming promoted CM proliferation by alleviating ROS and inducing M2 macrophage polarization, indicating that LDHA might be an effective target for promoting cardiac repair post-MI.


Assuntos
Infarto do Miocárdio , Miócitos Cardíacos , Animais , Proliferação de Células , Coenzima A/farmacologia , Lactato Desidrogenase 5 , Lactatos/metabolismo , Lactatos/farmacologia , Macrófagos/metabolismo , Mamíferos , Camundongos , Camundongos Knockout , Infarto do Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Tiorredoxina Redutase 1/metabolismo
11.
Obesity (Silver Spring) ; 30(10): 1995-2004, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36062886

RESUMO

OBJECTIVE: The study goal was to analyze the effects of a high-fat diet (HFD) on the histone 3 lysine 27 (H3K27) posttranscriptional modifications and the expression of histone-modifying enzymes in adipose-derived stromal cells (ASCs) from white adipose tissue (WAT). METHODS: Male C57BL/6J mice received control or HFD for 12 weeks. The ASCs were isolated from subcutaneous and visceral (epididymal) WAT, cultivated, and evaluated for expression of H3K27 trimethylation (H3K27me3) and H3K27 acetylation (H3K27ac) by Western blot. The transcription of histone-modifying enzymes was analyzed by real-time polymerase chain reaction. RESULTS: When compared with control, HFD ASCs showed a decrease in H3K27ac enrichment in subcutaneous and visceral WAT and ATP-citrate lyase expression in subcutaneous WAT. Curiously, the expression of CREB-binding protein was increased in visceral ASCs from HFD-fed mice. CONCLUSIONS: These results show that an HFD significantly reduces acetylation of H3K27 in ASCs and the expression of ATP-citrate lyase in subcutaneous ASCs, suggesting that, in this fat depot, the H3K27ac reduction could be partly due to lower acetyl-coenzyme A availability. H3K27ac is an epigenetic mark responsible for increasing the transcription rate and its reduction can have an important impact on ASC proliferation and differentiation potential.


Assuntos
Dieta Hiperlipídica , Histonas , Acetilação , Trifosfato de Adenosina , Animais , Proteína de Ligação a CREB/metabolismo , Coenzima A/metabolismo , Histonas/metabolismo , Lisina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células Estromais/metabolismo
12.
Biomolecules ; 12(9)2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36139163

RESUMO

Coenzyme A (CoA) is an essential co-factor at the intersection of diverse metabolic pathways. Cellular CoA biosynthesis is regulated at the first committed step-phosphorylation of pantothenic acid-catalyzed by pantothenate kinases (PANK1,2,3 in humans, PANK3 being the most highly expressed). Despite the critical importance of CoA in metabolism, the differential roles of PANK isoforms remain poorly understood. Our investigations of PANK proteins as potential precision oncology collateral lethality targets (PANK1 is co-deleted as part of the PTEN locus in some highly aggressive cancers) were severely hindered by a dearth of commercial antibodies that can reliably detect endogenous PANK3 protein. While we successfully validated commercial antibodies for PANK1 and PANK2 using CRISPR knockout cell lines, we found no commercial antibody that could detect endogenous PANK3. We therefore set out to generate a mouse monoclonal antibody against human PANK3 protein. We demonstrate that a clone (Clone MDA-299-62A) can reliably detect endogenous PANK3 protein in cancer cell lines, with band-specificity confirmed by CRISPR PANK3 knockout and knockdown cell lines. Sub-cellular fractionation shows that PANK3 is overwhelmingly cytosolic and expressed broadly across cancer cell lines. PANK3 monoclonal antibody MDA-299-62A should prove a valuable tool for researchers investigating this understudied family of metabolic enzymes in health and disease.


Assuntos
Neoplasias , Ácido Pantotênico , Animais , Anticorpos Monoclonais , Coenzima A , Humanos , Camundongos , Medicina de Precisão , Isoformas de Proteínas
13.
Genes (Basel) ; 13(9)2022 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-36140759

RESUMO

The tree peony (Paeonia ostii 'Feng Dan Bai') has excellent drought tolerance. Although it has already been reported that the cuticle is an essential barrier against drought stress, the critical genes for cuticle resistance to drought remain unclear. However, the long-chain acyl-CoA synthetases (LACS) family of genes may be significant for the synthesis of cuticle wax. To test whether the LACS gene family is involved in cuticle response to drought stress in tree peony, we measure the thickness of cuticle stems and leaves alongside LACS enzyme activity. It is found that the cuticle thickens and the LACS enzyme increases with the maturation of stems and leaves, and there is a positive correlation between them. The LACS enzyme increases within 12 h under drought stress induced by polyethylene glycol (PEG). The transcriptome sequencing result (BioProject accession number PRJNA317164) is searched for, and a LACS gene with high expression is cloned. This gene has high homology and similarity with LACS4 from Arabidopsis thaliana. The gene is named PoLACS4. It is show to be highly expressed in mature leaves and peaks within 1 h under drought and salt stresses. All these results suggest that the LACS family of genes may be involved in cuticle response to drought stress and that PoLACS4 is a crucial gene which responds rapidly to drought in the tree peony.


Assuntos
Arabidopsis , Paeonia , Arabidopsis/genética , Coenzima A/genética , Coenzima A/metabolismo , Coenzima A Ligases/genética , Secas , Regulação da Expressão Gênica de Plantas , Paeonia/genética , Polietilenoglicóis
14.
PLoS One ; 17(9): e0272986, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36054206

RESUMO

Acyl-CoA synthetase 1 (ACSL1) is an enzyme that converts fatty acids to acyl-CoA-derivatives for lipid catabolism and lipid synthesis in general and can provide substrates for the production of mediators of inflammation in monocytes and macrophages. Acsl1 expression is increased by hyperglycemia and inflammatory stimuli in monocytes and macrophages, and promotes the pro-atherosclerotic effects of diabetes in mice. Yet, surprisingly little is known about the mechanisms underlying Acsl1 transcriptional regulation. Here we demonstrate that the glucose-sensing transcription factor, Carbohydrate Response Element Binding Protein (CHREBP), is a regulator of the expression of Acsl1 mRNA by high glucose in mouse bone marrow-derived macrophages (BMDMs). In addition, we show that inflammatory stimulation of BMDMs with lipopolysaccharide (LPS) increases Acsl1 mRNA via the transcription factor, NF-kappa B. LPS treatment also increases ACSL1 protein abundance and localization to membranes where it can exert its activity. Using an Acsl1 reporter gene containing the promoter and an upstream regulatory region, which has multiple conserved CHREBP and NF-kappa B (p65/RELA) binding sites, we found increased Acsl1 promoter activity upon CHREBP and p65/RELA expression. We also show that CHREBP and p65/RELA occupy the Acsl1 promoter in BMDMs. In primary human monocytes cultured in high glucose versus normal glucose, ACSL1 mRNA expression was elevated by high glucose and further enhanced by LPS treatment. Our findings demonstrate that CHREBP and NF-kappa B control Acsl1 expression under hyperglycemic and inflammatory conditions.


Assuntos
Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Coenzima A Ligases/genética , Hiperglicemia , Inflamação/metabolismo , Subunidade p50 de NF-kappa B/metabolismo , NF-kappa B , Animais , Coenzima A/metabolismo , Glucose/metabolismo , Glucose/farmacologia , Humanos , Hiperglicemia/genética , Hiperglicemia/metabolismo , Inflamação/genética , Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Camundongos , NF-kappa B/metabolismo , RNA Mensageiro/genética
15.
Microb Genom ; 8(9)2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36125959

RESUMO

There is an urgent need to replace petroleum-based plastic with bio-based and biodegradable alternatives. Polyhydroxyalkanoates (PHAs) are attractive prospective replacements that exhibit desirable mechanical properties and are recyclable and biodegradable in terrestrial and marine environments. However, the production costs today still limit the economic sustainability of the PHA industry. Seaweed cultivation represents an opportunity for carbon capture, while also supplying a sustainable photosynthetic feedstock for PHA production. We mined existing gene and protein databases to identify bacteria able to grow and produce PHAs using seaweed-derived carbohydrates as substrates. There were no significant relationships between the genes involved in the deconstruction of algae polysaccharides and PHA production, with poor to negative correlations and diffused clustering suggesting evolutionary compartmentalism. We identified 2 987 bacterial candidates spanning 40 taxonomic families predominantly within Alphaproteobacteria, Gammaproteobacteria and Burkholderiales with enriched seaweed-degrading capacity that also harbour PHA synthesis potential. These included highly promising candidates with specialist and generalist specificities, including Alteromonas, Aquisphaera, Azotobacter, Bacillus, Caulobacter, Cellvibrionaceae, Duganella, Janthinobacterium, Massilia, Oxalobacteraceae, Parvularcula, Pirellulaceae, Pseudomonas, Rhizobacter, Rhodanobacter, Simiduia, Sphingobium, Sphingomonadaceae, Sphingomonas, Stieleria, Vibrio and Xanthomonas. In this enriched subset, the family-level densities of genes targeting green macroalgae polysaccharides were considerably higher (n=231.6±68.5) than enzymes targeting brown (n=65.34±13.12) and red (n=30.5±10.72) polysaccharides. Within these organisms, an abundance of FabG genes was observed, suggesting that the fatty acid de novo synthesis pathway supplies (R)-3-hydroxyacyl-CoA or 3-hydroxybutyryl-CoA from core metabolic processes and is the predominant mechanism of PHA production in these organisms. Our results facilitate extending seaweed biomass valorization in the context of consolidated biorefining for the production of bioplastics.


Assuntos
Petróleo , Poli-Hidroxialcanoatos , Alga Marinha , Bactérias/genética , Bactérias/metabolismo , Carboidratos , Carbono/metabolismo , Coenzima A/metabolismo , Ácidos Graxos/metabolismo , Humanos , Petróleo/metabolismo , Plásticos/metabolismo , Poli-Hidroxialcanoatos/química , Poli-Hidroxialcanoatos/metabolismo , Estudos Prospectivos
16.
Int J Mol Sci ; 23(18)2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36142850

RESUMO

In this study, we identified a novel glossy mutant from Chinese cabbage, named SD369, and all wax monomers longer than 26 carbons were significantly decreased. Inheritance analysis revealed that the glossy trait of SD369 was controlled by a single recessive locus, BrWAX3. We fine-mapped the BrWAX3 locus to an interval of 161.82 kb on chromosome A09. According to the annotated genome of Brassica rapa, Bra024749 (BrCER60.A09), encoding a ß-ketoacyl-CoA synthase, was identified as the candidate gene. Expression analysis showed that BrCER60.A09 was significantly downregulated in all aerial organs of glossy plants. Subcellular localization indicated that the BrCER60.A09 protein functions in the endoplasmic reticulum. A 5567-bp insertion was identified in exon 1 of BrCER60.A09 in SD369, which lead to a premature stop codon, thus causing a loss of function of the BrCER60.A09 enzyme. Moreover, comparative transcriptome analysis revealed that the 'cutin, suberine, and wax biosynthesis' pathway was significantly enriched, and genes involved in this pathway were almost upregulated in glossy plants. Further, two functional markers, BrWAX3-InDel and BrWAX3-KASP1, were developed and validated. Overall, these results provide a new information for the cuticular wax biosynthesis and provide applicable markers for marker-assisted selection (MAS)-based breeding of Brassica rapa.


Assuntos
Brassica rapa , Brassica , Brassica/genética , Brassica/metabolismo , Brassica rapa/genética , Brassica rapa/metabolismo , China , Códon sem Sentido/metabolismo , Coenzima A/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Ceras/metabolismo
17.
Proc Natl Acad Sci U S A ; 119(39): e2208496119, 2022 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-36122204

RESUMO

Allotetraploid cotton (Gossypium) species represents a model system for the study of plant polyploidy, molecular evolution, and domestication. Here, chromosome-scale genome sequences were obtained and assembled for two recently described wild species of tetraploid cotton, Gossypium ekmanianum [(AD)6, Ge] and Gossypium stephensii [(AD)7, Gs], and one early form of domesticated Gossypium hirsutum, race punctatum [(AD)1, Ghp]. Based on phylogenomic analysis, we provide a dated whole-genome level perspective for the evolution of the tetraploid Gossypium clade and resolved the evolutionary relationships of Gs, Ge, and domesticated G. hirsutum. We describe genomic structural variation that arose during Gossypium evolution and describe its correlates-including phenotypic differentiation, genetic isolation, and genetic convergence-that contributed to cotton biodiversity and cotton domestication. Presence/absence variation is prominent in causing cotton genomic structural variations. A presence/absence variation-derived gene encoding a phosphopeptide-binding protein is implicated in increasing fiber length during cotton domestication. The relatively unimproved Ghp offers the potential for gene discovery related to adaptation to environmental challenges. Expanded gene families enoyl-CoA δ isomerase 3 and RAP2-7 may have contributed to abiotic stress tolerance, possibly by targeting plant hormone-associated biochemical pathways. Our results generate a genomic context for a better understanding of cotton evolution and for agriculture.


Assuntos
Fibra de Algodão , Gossypium , Coenzima A , Gossypium/genética , Isomerases , Fosfopeptídeos , Reguladores de Crescimento de Plantas , Tetraploidia
18.
Medicina (Kaunas) ; 58(9)2022 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-36143922

RESUMO

Background and Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) isolates are associated with various diseases ranged from mild superficial impairments to invasive infections. This study aimed to evaluate the ability of polymerase chain reaction (PCR) based methods namely, restriction fragment length polymorphism (RFLP) of the coa gene and random amplified polymorphic DNA (RAPD), to determine the genetic diversity of MRSA isolates. Materials and Methods: A total of 37 MRSA isolates were conventionally identified depending on their biochemical and microbiological culture characteristics. Genotypic confirmation was based on detection of the associated mecA gene. The genetic variation amongst MRSA isolates was evaluated following the coa gene-based RFLP and RAPD fingerprints. Results: Results illustrated that, the species specific coa gene was detected in all MRSA isolates. The irregular bands intensity, number, and molecular sizes of the PCR amplicons demonstrated the coa gene polymorphism. The incompatible AluI digestion patterns of these amplicons classified the tested MRSA isolates into 20 RFLP patterns which confirm the coa gene polymorphism. Additionally, the PCR-based RAPD analysis showed variable bands number with size range of approximately 130 bp to 4 kbp, which indicated the genetic variation of the tested MRSA isolates as it created 36 variable RAPD banding profiles. Conclusions: coa gene AluI enzymatic restriction sites, amongst the tested MRSA isolates, certify their genetic variation on the basis of the accurate but complicated and relatively expensive coa gene-based RFLP. Conversely, the results verified the excellent ability of the simple and cost-effective PCR-based RAPD analysis to discriminate between MRSA isolates without any preface data about the genome.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Coagulase/genética , Coagulase/farmacologia , Coenzima A/genética , Coenzima A/farmacologia , DNA/farmacologia , Humanos , Staphylococcus aureus Resistente à Meticilina/genética , Polimorfismo de Fragmento de Restrição , Técnica de Amplificação ao Acaso de DNA Polimórfico , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus/genética
19.
Nat Commun ; 13(1): 5696, 2022 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-36171419

RESUMO

Fatty liver is a highly heterogenous condition driven by various pathogenic factors in addition to the severity of steatosis. Protein insufficiency has been causally linked to fatty liver with incompletely defined mechanisms. Here we report that fatty liver is a sulfur amino acid insufficient state that promotes metabolic inflexibility via limiting coenzyme A availability. We demonstrate that the nutrient-sensing transcriptional factor EB synergistically stimulates lysosome proteolysis and methionine adenosyltransferase to increase cysteine pool that drives the production of coenzyme A and glutathione, which support metabolic adaptation and antioxidant defense during increased lipid influx. Intriguingly, mice consuming an isocaloric protein-deficient Western diet exhibit selective hepatic cysteine, coenzyme A and glutathione deficiency and acylcarnitine accumulation, which are reversed by cystine supplementation without normalizing dietary protein intake. These findings support a pathogenic link of dysregulated sulfur amino acid metabolism to metabolic inflexibility that underlies both overnutrition and protein malnutrition-associated fatty liver development.


Assuntos
Aminoácidos Sulfúricos , Fígado Gorduroso , Aminoácidos Sulfúricos/metabolismo , Animais , Antioxidantes/metabolismo , Coenzima A/metabolismo , Cisteína/metabolismo , Cistina/metabolismo , Proteínas na Dieta/metabolismo , Fígado Gorduroso/metabolismo , Glutationa/metabolismo , Homeostase , Lipídeos , Fígado/metabolismo , Metionina/metabolismo , Metionina Adenosiltransferase/metabolismo , Camundongos , Oxirredução
20.
BMC Microbiol ; 22(1): 227, 2022 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-36171563

RESUMO

BACKGROUND: Environmental contamination from synthetic plastics and their additives is a widespread problem. Phthalate esters are a class of refractory synthetic organic compounds which are widely used in plastics, coatings, and for several industrial applications such as packaging, pharmaceuticals, and/or paints. They are released into the environment during production, use and disposal, and some of them are potential mutagens and carcinogens. Isophthalate (1,3-benzenedicarboxylic acid) is a synthetic chemical that is globally produced at a million-ton scale for industrial applications and is considered a priority pollutant. Here we describe the biochemical characterization of an enzyme involved in anaerobic degradation of isophthalate by the syntrophically fermenting bacterium Syntrophorhabdus aromaticivorans strain UI that activate isophthalate to isophthalyl-CoA followed by its decarboxylation to benzoyl-CoA. RESULTS: Isophthalate:Coenzyme A ligase (IPCL, AMP-forming) that activates isophthalate to isophthalyl-CoA was heterologously expressed in E. coli (49.6 kDa) for biochemical characterization. IPCL is homologous to phenylacetate-CoA ligase that belongs to the family of ligases that form carbon-sulfur bonds. In the presence of coenzyme A, Mg2+ and ATP, IPCL converts isophthalate to isophthalyl-CoA, AMP and pyrophosphate (PPi). The enzyme was specifically induced after anaerobic growth of S. aromaticivorans in a medium containing isophthalate as the sole carbon source. Therefore, IPCL exhibited high substrate specificity and affinity towards isophthalate. Only substrates that are structurally related to isophthalate, such as glutarate and 3-hydroxybenzoate, could be partially converted to the respective coenzyme A esters. Notably, no activity could be measured with substrates such as phthalate, terephthalate and benzoate. Acetyl-CoA or succinyl-CoA did not serve as CoA donors. The enzyme has a theoretical pI of 6.8 and exhibited optimal activity between pH 7.0 to 7.5. The optimal temperature was between 25 °C and 37 °C. Denaturation temperature (Tm) of IPCL was found to be at about 63 °C. The apparent KM values for isophthalate, CoA, and ATP were 409 µM, 642 µM, and 3580 µM, respectively. Although S. aromaticivorans is a strictly anaerobic bacterium, the enzyme was found to be oxygen-insensitive and catalysed isophthalyl-CoA formation under both anoxic and oxic conditions. CONCLUSION: We have successfully cloned the ipcl gene, expressed and characterized the corresponding IPCL enzyme, which plays a key role in isophthalate activation that initiates its activation and further degradation by S. aromaticivorans. Its biochemical characterization represents an important step in the elucidation of the complete degradation pathway of isophthalate.


Assuntos
Difosfatos , Poluentes Ambientais , Acetilcoenzima A/metabolismo , Monofosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Anaerobiose , Composição de Bases , Benzoatos/metabolismo , Carbono , Carcinógenos , Coenzima A/metabolismo , Coenzima A Ligases , Escherichia coli/metabolismo , Glutaratos , Hidroxibenzoatos , Mutagênicos , Oxigênio , Fenilacetatos/metabolismo , Ácidos Ftálicos , Filogenia , Plásticos , RNA Ribossômico 16S , Análise de Sequência de DNA , Enxofre , Xenobióticos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...