Adenosine Triphosphate Synthase Research Articles

Acetate ameliorates ovarian mitochondrial dysfunction in letrozole-induced polycystic ovarian syndrome rat model by improving mitofusin-2

Androgen excess and metabolic abnormality largely contribute to the pathogenesis of polycystic ovarian syndrome (PCOS), which primarily precipitates ovarian dysfunction and infertility in reproductive-age women. Impaired mitochondrial function and epigenetic alteration have been linked to the development of PCOS. However, it is unknown whether acetate would exert a therapeutic effect on ovarian mitochondrial dysfunction in PCOS. Herein, the study hypothesized that acetate reverses ovarian mitochondrial dysfunction in experimental PCOS rat model, possibly through modulation of mitofusin-2 (MFn2). Eight-week-old female Wistar rats were randomized into four groups (n = 5). Induction of PCOS was performed by 1 mg/kg letrozole (p.o.), administered for 21 days. Thereafter, the rats were treated with acetate (200 mg/kg; p.o.) for 6 weeks. The PCOS rats demonstrated androgen excess, multiple ovarian cysts, elevated anti-mullerian hormone and leptin and decreased SHBG, adiponectin and 17-β estradiol with corresponding increase in ovarian transforming growth factor-β1. Additionally, inflammation (tumor growth factor and nuclear factor-kB), elevated caspase-6, decreased hypoxia-inducible factor-1α and elevated histone deacetylase-2 (HDAC2) were observed in the ovaries of PCOS rats, while mitochondrial abnormality with evidence of decreased adenosine triphosphate synthase and MFn2 was observed in rats with PCOS. Treatment with acetate reversed the alterations. The present results collectively suggest that acetate ameliorates ovarian mitochondrial abnormality, a beneficial effect that is accompanied by MFn2 with consequent normalization of reproductive-endocrine profile and ovarian function. Perhaps, the present data provide hope for PCOS individuals that suffer infertility.

Method Development and Validation for Estimation of Bedaquiline and in Tablet Dosage form by Hptlc and Rp-Hplc Study

A diarylquinoline anti-mycobacterial drug, bedaquiline (BDQ), treats mycobacterial infections. It inhibits the proton pump by inhibiting mycobacterial Adenosine Triphosphate (ATP) synthase. ATP enzyme that is energy production requires Mycobacterium tuberculosis. The present study aimed to develop three analytical methods, i.e., High-performance thin-layer chromatography (HPTLC), HPLC, and UV, as BDQ estimation in tablet forms. HPTLC is employed in the first method, which uses ethanol-chloroform-toluene as the mobile phase (6:3.5:0.5). UV spectroscopy at 240nm was used to analyse BDQ at 100-300 micrograms per milliliter. RP-HPLC method based on the separation using a C18 column, Buffer (sodium acetate): Methyl alcohol (70:30) as mobile phase flowing at a rate of 1ml per min. The retention time was determined to be 5.4 minutes. At 240 nm, it could quantify the concentrations of 5-25 g per ml at concentrations of 5-20 g per ml. The third method involves the "UV-VISIBLE” spectrophotometric method. A range of BDQ concentrations is determined of 8-40µg/ml, respectively. These methods were checked to ICH guidelines, and as a result, a commercially available pharmaceutical formulation, with the findings of statistical comparison, was conducted between all three methods.

Mechanism of proton-powered c-ring rotation in a mitochondrial ATP synthase

Proton-powered c-ring rotation in mitochondrial ATP synthase is crucial to convert the transmembrane protonmotive force into torque to drive the synthesis of adenosine triphosphate (ATP). Capitalizing on recent cryo-EM structures, we aim at a structural and energetic understanding of how functional directional rotation is achieved. We performed multi-microsecond atomistic simulations to determine the free energy profiles along the c-ring rotation angle before and after the arrival of a new proton. Our results reveal that rotation proceeds by dynamic sliding of the ring over the a-subunit surface, during which interactions with conserved polar residues stabilize distinct intermediates. Ordered water chains line up for a Grotthuss-type proton transfer in one of these intermediates. After proton transfer, a high barrier prevents backward rotation and an overall drop in free energy favors forward rotation, ensuring the directionality of c-ring rotation required for the thermodynamically disfavored ATP synthesis. The essential arginine of the a-subunit stabilizes the rotated configuration through a salt bridge with the c-ring. Overall, we describe a complete mechanism for the rotation step of the ATP synthase rotor, thereby illuminating a process critical to all life at atomic resolution.

Mitochondrial Dysfunction in Advanced Maternal Aged Cumulus Cells: A Possible Link to ATP Synthase Impairment?

Age-related changes in the mitochondrial status of human cumulus cells (hCCs) impact oocyte quality; however, the relationship between hCC mitochondrial (dys)function and reproductive aging remains poorly understood. This study aimed to establish the interplay between hCC mitochondrial dysfunction and women's reproductive potential. In this investigation, 266 women were enrolled and categorized into two groups based on their age: a young group (<35 years old) and an advanced maternal age (AMA) group (≥35 years old). Comprehensive analysis of reproductive outcomes was conducted in our population. Various mitochondrial-related parameters were analyzed across distinct subsets. Specifically, mitochondrial membrane potential (∆Ψm) and mitochondrial mass were examined in 53 samples, mtDNA content in 25 samples, protein levels in 23 samples, bioenergetic profiles using an XF24 Extracellular Flux Analyzer in 6 samples, and levels of reactive oxygen species (ROS) and adenosine triphosphate (ATP) in 39 and 43 samples, respectively. In our study, the reproductive potential of AMA women sharply decreased, as expected. Additionally, an impairment in the mitochondrial function of hCCs in older women was observed; however, no differences were found in terms of mitochondrial content. Regarding oxidative phosphorylation, metabolic profiling of hCCs from AMA women indicated a decrease in respiratory capacity, which was correlated with an age-dependent decrease in the ATP synthase (ATP5A1) protein level. However, intracellular ROS and ATP levels did not differ between groups. In conclusion, our study indicates that age-related dysfunction in hCCs is associated with impaired mitochondrial function, and, although further studies are required, ATP synthase could be relevant in this impairment.

Conformational ensemble of yeast ATP synthase at low pH reveals unique intermediates and plasticity in F1-Fo coupling.

Mitochondrial adenosine triphosphate (ATP) synthase uses the proton gradient across the inner mitochondrial membrane to synthesize ATP. Structural and single molecule studies conducted mostly at neutral or basic pH have provided details of the reaction mechanism of ATP synthesis. However, pH of the mitochondrial matrix is slightly acidic during hypoxia and pH-dependent conformational changes in the ATP synthase have been reported. Here we use single-particle cryo-EM to analyze the conformational ensemble of the yeast (Saccharomyces cerevisiae) ATP synthase at pH 6. Of the four conformations resolved in this study, three are reaction intermediates. In addition to canonical catalytic dwell and binding dwell structures, we identify two unique conformations with nearly identical positions of the central rotor but different catalytic site conformations. These structures provide new insights into the catalytic mechanism of the ATP synthase and highlight elastic coupling between the catalytic and proton translocating domains.

Insilico exploration C. koseri ATP synthase inhibitors by pharmacophore-based virtual screening, molecular docking and MD simulation.

Citrobacter koseri is a gram-negative rod that causes infections in people who have significant comorbidities and are immunocompromised. Antibiotic-resistant strains are becoming more common, which complicates infection treatment and highlights the need for innovative, effective drugs to fight these resistant strains. The enzyme complex ATP synthase participates in the adenosine triphosphate (ATP) synthesis, the fundamental energy currency of cells. This study used Computer-Aided Drug Design approaches to identify potential inhibitors of C. koseri ATP synthase. SWISS-MODEL was used to predict the 3D structure of C. koseri ATP synthase. A ligand-based pharmacophore model was developed using chemical features of ampicillin. Following ligand-based virtual screening across nine databases, the 2043 screened hits were docked to the ATP synthase active site using the standard precision mode of the glide tool. Based on their binding affinities, the top ten compounds were selected for additional investigation. The binding affinities of the chosen compounds ranged from -10.021 to -8.452 kcal/mol. The top four compounds (PubChem-25230613, PubChem-74936833, CHEMBL263035, PubChem-44208924) with the best ADMET characteristics and binding modes were chosen. Thus, the feasible binding mechanisms of the selected compounds were subjected to stability analysis using the MD Simulation study, which revealed the compounds' stability as potent inhibitors within the protein binding pocket. This computational approach provides important insights into the rational design of novel therapeutics and emphasizes the importance of targeting essential metabolic pathways when combating antibiotic-resistant pathogens. Future experimental validation and optimization of the identified inhibitors is required to determine their efficacy and safety profiles for clinical use.

A simple assay for inhibitors of mycobacterial oxidative phosphorylation

Oxidative phosphorylation, the combined activities of the electron transport chain (ETC) and adenosine triphosphate (ATP) synthase, has emerged as a valuable target for antibiotics to treat infection with Mycobacterium tuberculosis and related pathogens. In oxidative phosphorylation, the ETC establishes a transmembrane electrochemical proton gradient that powers ATP synthesis. Monitoring oxidative phosphorylation with luciferase-based detection of ATP synthesis or measurement of oxygen consumption can be technically challenging and expensive. These limitations reduce the utility of these methods for characterization of mycobacterial oxidative phosphorylation inhibitors. Here we show that fluorescence-based measurement of acidification of inverted membrane vesicles (IMVs) can detect and distinguish between inhibition of the ETC, inhibition of ATP synthase, and non-specific membrane uncoupling. In this assay, IMVs from M. smegmatis are acidified either through the activity of the ETC or ATP synthase, the latter modified genetically to allow it to serve as an ATP-driven proton pump. Acidification is monitored by fluorescence from 9-amino-6-chloro-2-methoxyacridine, which accumulates and quenches in acidified IMVs. Non-specific membrane uncouplers prevent both succinate- and ATP-driven IMV acidification. In contrast, the ETC Complex III2IV2 inhibitor telacebec (Q203) prevents succinate-driven acidification but not ATP-driven acidification and the ATP synthase inhibitor bedaquiline prevents ATP-driven acidification but not succinate-driven acidification. We use the assay to show that, as proposed previously, lansoprazole sulfide is an inhibitor of Complex III2IV2 while thioridazine uncouples the mycobacterial membrane non-specifically. Overall, the assay is simple, low cost, and scalable, which will make it useful for identifying and characterizing new mycobacterial oxidative phosphorylation inhibitors.

The role of photorespiration in preventing feedback regulation via ATP synthase in Nicotiana tabacum.

Photorespiration consumes substantial amounts of energy in the forms of adenosine triphosphate (ATP) and reductant making the pathway an important component in leaf energetics. Because of this high reductant demand, photorespiration is proposed to act as a photoprotective electron sink. However, photorespiration consumes more ATP relative to reductant than the C3 cycle meaning increased flux disproportionally increases ATP demand relative to reductant. Here we explore how energetic consumption from photorespiration impacts the flexibility of the light reactions in nicotiana tabacum. Specifically, we demonstrate that decreased photosynthetic efficiency (ϕII ) at low photorespiratory flux was related to feedback regulation at the chloroplast ATP synthase. Additionally, decreased ϕII at high photorespiratory flux resulted in the accumulation of photoinhibition at photosystem II centers. These results are contrary to the proposed role of photorespiration as a photoprotective electron sink. Instead, our results suggest a novel role of ATP consumption from photorespiration in maintaining ATP synthase activity, with implications for maintaining energy balance and preventing photodamage that will be critical for plant engineering strategies.

Interaction between mitochondrial homeostasis and barrier function in lipopolysaccharide-induced endothelial cell injury.

This study aimed to investigate the effects of mitochondrial homeostasis on lipopolysaccharide (LPS)-induced endothelial cell barrier function and the mechanisms that underlie these effects. Cells were treated with LPS or oligomycin (mitochondrial adenosine triphosphate synthase inhibitor) and the mitochondrial morphology, mitochondrial reactive oxygen species (mtROS), and mitochondrial membrane potential (ΔΨm) were evaluated. Moreover, the shedding of glycocalyx-heparan sulphate (HS), the levels of HS-specific degrading enzyme heparanase (HPA), and the expression of occludin and zonula occludens (ZO-1) of Tight Junctions (TJ)s, which are mediated by myosin light chain phosphorylation (p-MLC), were assessed. Examining the changes in mitochondrial homeostasis showed that adding heparinase III, which is an exogenous HPA, can destroy the integrity of glycocalyx. LPS simultaneously increased mitochondrial swelling, mtROS, and ΔΨm. Without oligomycin effects, HS, HPA levels, and p-MLC were found to be elevated, and the destruction of occludin and ZO-1 increased. Heparinase III not only damaged the glycocalyx by increasing HS shedding but also increased mitochondrial swelling and mtROS and decreased ΔΨm. Mitochondrial homeostasis is involved in LPS-induced endothelial cell barrier dysfunction by aggravating HPA and p-MLC levels. In turn, the integrated glycocalyx protects mitochondrial homeostasis.

Dangua Fang regulating tricarboxylic acid cycle and respiratory chain and its mechanism in diabetic rats.

To investigate the influence and possible targets of Dangua Fang on tricarboxylic acid (TCA) cycle and respiratory chain to enrich the prescription's mechanism of effective intervention on glycolipid metabolic diseases such as type 2 diabetes. After interventional rats were fed with high glucose and high fat diet ad libitum for 4 weeks, intraperitoneally injected streptozotocin to induce diabetic model. According to blood glucose level,28 diabetic rats were selected and continued to be fed with high glucose and high fat diet, were stratified by body weight, and divided randomly by blood glucose into Model group (was given sterile water by gastric perfusion and injected aquae pro injection intraperitoneally), Dangua group [Dangua liquor 20.5 g·kg-1·d-1 by perfusion and aquae pro injection intraperitoneally], Inhibitor group [sterile water by perfusion and nicotinamide phosphoribosyl transferase (Nampt) specific blocker GEN-617 1.25 mg/kg intraperitoneally], DanInhit group (Dangua liquor and GEN-617 synchronously). Control group were continuously fed with ordinary diet. The intervention was last for 10 weeks. Body weight (BW), liver index (LI), glycosylated hemoglobin (HbA1c), TC, TG, free fatty acids (FFA), creatinine (Cr), and A-ketoglutarate (α-KG), Iso-citric acid (ICA), oxaloacetic acid (OAA) were tested. The cytochrome C oxidase (COX) and Succinate dehydrogenase (SDH) were evaluated by Colorimetry; Nampt protein, Adenosine triphosphate (ATP) synthase (ATPs), Nicotinamide adenine dinucleotide (NAD+)and its reduced (NADH) in liver were measured by enzyme linked immunosorbent assay. The expressions of Nampt and mitochondrialnadhdehydrogenase-1 (mt-ND1) gene in liver was assessed by real-time polymerase chain reaction. Hepatic tissue staining was also completed. The levels of BW, ICA, α-KG and Nampt-mRNA in the Model group are lower than that in the Normal group (P < 0.05), conversely, liver weight, LI, TC, HbA1c, SDH and ATPs, mt-ND1-mRNA, and Nampt protein in the Model group are higher (P < 0.01, P < 0.05). Compared with Model group, the levels of ICA, Nampt-mRNA and Nampt in Dangua group are significantly increased, and FFA obviously raised (P < 0.01 and P < 0.05); liver weight, BW, SDH are obviously lower, and HbA1c decreased significantly (P < 0.01, P < 0.05). TG, FFA and Nampt protein increased in the DanInhit group, TC, TG, BW obviously increased in the Inhibitor group, but SDH is decreased in both the two groups (P < 0.05, P < 0.01). Compared with Dangua group, DanInhib group has the lower levels of ICA, mt-ND1-mRNA, Nampt-mRNA, and the higher level of BW, LI and HbA1c. In the Inhibitor group, ICA and Nampt protein decreased, BW and LI, HbA1c and TG increased (P < 0.01 or P < 0.05). Tissue staining display that, in the model group there is obvious pathologic changes ie: fibrosis, steatosis and inflammatory cell infiltration. Lesions in the Dangua group are mild, and those of Inhibitor group are more obvious than the Model group, and DanInhit group is intermediately affected compared to Dangua group and Inhibitor group. Dangua Fang increases the metabolic flux of TCA cycle and optimizes respiratory chain function by up-regulating Nampt expression.

Effects of Curcumin Treatment on Cell Energy Status, Levels of Mitochondrial Enzymes, and Gene Expression of Glucose-related Mechanism in Pancreatic Cancer Cell Lines

Background and Purpose:. Curcumin is an active component of turmeric, has antitumor, immunomodulatory, anti-inflammatory effects. It was aimed to investigate the effects of the administration of curcumin on the energy metabolism, the abnormal redox defense mechanism profile, the malignant transformation indicator of Panc-1 and BxPC-3 pancreatic cancer cells. &#x0D; Methods: BxPC-3 and Panc-1 cells were incubated, were replaced with containing various concentrations of curcumin (10-125 μM) for 24 h. Cell lysate Adenosine triphosphate (ATP), Adenosine diphosphate (ADP), Adenosine monophosphate (AMP), Manganese superoxidase (MnSOD), and cytochrome p450 reductase (CPR) concentrations were analyzed with HPLC and ELISA methods. Genes expression of Lactate dehydrogenase (LDH), mitochondrially encoded ATP synthase membrane subunit 6 (MTATP6), Glucose transporter 1 (GLUT1), and cytochrome p450 were analyzed. &#x0D; Results and Conclusion: IC50 values for 24 hours were found as 47,26 μM in BxPC-3 and 45,84 μM in Panc-1 cells. Treatment with curcumin inhibits oxidative stress by increasing MnSOD enzyme levels. ATP levels did not change in BxPC-3 cells, but it showed an increase in Panc-1 supplemented with curcumin. The effects of curcumin on GLUT-1 are significantly important at a dose of curcumin of 45 μM concentration and affect glucose consumption in both cells. Curcumin showed anti-proliferative, and antioxidant effects.

Hepatic ischemia-reperfusion syndrome and its effect on the cardiovascular system: The role of treprostinil, a synthetic prostacyclin analog.

Hepatic ischemia-reperfusion syndrome has been the subject of intensive study and experimentation in recent decades since it is responsible for the outcome of several clinical entities, such as major hepatic resections and liver transplantation. In addition to the organ's post reperfusion injury, this syndrome appears to play a central role in the dysfunction of distant tissues and systems. Thus, continuous research should be directed toward finding effective therapeutic options to improve the outcome and reduce the postoperative morbidity and mortality rates. Treprostinil is a synthetic analog of prostaglandin I2, and its experimental administration has shown encouraging results. It has already been approved by the Food and Drug Administration in the United States for pulmonary arterial hypertension and has been used in liver transplantation, where preliminary encouraging results showed its safety and feasibility by using continuous intravenous administration at a dose of 5 ng/kg/min. Treprostinil improves renal and hepatic function, diminishes hepatic oxidative stress and lipid peroxidation, reduces hepatictoll-like receptor 9 and inflammation, inhibits hepatic apoptosis and restores hepatic adenosine triphosphate (ATP) levels and ATP synthases, which is necessary for functional maintenance of mitochondria. Treprostinil exhibits vasodilatory properties and antiplatelet activity and regulates proinflammatory cytokines; therefore, it can potentially minimize ischemia-reperfusion injury. Additionally, it may have beneficial effects on cardiovascular parameters, and much current research interest is concentrated on this compound.

Unraveling gene expression and genetic instability in dental fluorosis: Investigating the impact of chronic fluoride exposure

Chronic fluoride exposure, even in small quantities, when continuously ingested by the human population, can lead to a significant public health concern known as fluorosis. Our understanding of the effects of fluoride on human health, as well as its potential to impact DNA, is limited. The present study aimed to assess genetic instability in 20 individuals diagnosed with dental fluorosis and 20 individuals without the condition from the state of Rio Grande do Sul, Brazil. The participants' dental fluorosis was evaluated using the Thylstrup-Fejerskov index (TF). To further evaluate genetic instability, several assays were conducted, including the alkaline and modified (+FPG) comet assay (using a visual score, VS), the buccal micronucleus (MN) cytome (BMCyt) assay, the cytokinesis-block MN (CBMN-Cyt) assay, and the measurement of telomere length (TL). In addition, the study utilized tools from Systems Biology to gain insights into the effects of fluoride exposure on humans, which aided in the selection and evaluation of mRNA expression levels of specific genes, namely PPA1 (inorganic pyrophosphatase 1), AQP5 (Aquaporin 5), and MT-ATP6 (Mitochondrially Encoded Adenosine Triphosphate Synthase Membrane Subunit 6). Furthermore, fluoride levels in the blood and urine were assessed using an ion-selective electrode, along with the evaluation of the inflammatory response in serum. The group with dental fluorosis exhibited 2.18 times higher MN frequencies specifically when assessed using the CBMN-Cyt assay, in comparison with individuals without fluorosis. Findings from the enzyme-modified comet assay indicated oxidative damage to purines in DNA. Furthermore, a decrease in TL was observed, along with elevated expression patterns of the PPA1 and AQP5 genes, and significant alterations in cytokine release. Significant correlations were identified between the TF and age, as well as the levels of necrotic cells. Additionally, noteworthy correlations were established between fluoride levels and the levels of MN, VS, and MT-ATP6. Although dental fluorosis results from fluoride exposure, our research highlights the potential influence of this condition on genomic instability and gene expression. Consequently, our findings stress the importance of continuously monitoring populations with a high incidence of dental fluorosis to enhance our comprehension of how genomic instability might correlate with the origins and consequences of health problems in these individuals.

Cloning and functional analysis prohibitins protein-coding gene EuPHB1 in Eucommia ulmoides Oliver

As multifunctional proteins, prohibitins(PHBs) participate in many cellular processes and play essential roles in organisms. In this study, using rapid amplification of cDNA end (RACE) technology, EuPHB1 was cloned from Eucommia ulmoides Oliver (E. ulmoides). A subcellular localization assay preliminarily located EuPHB1 in mitochondria. Then EuPHB1 was transformed into tobacco, and phenotype analyses showed that overexpression of EuPHB1 caused leaves to become chlorotic and shrivel. Furthermore, genes related to hormone and auxin signal transduction, auxin binding, and transport, such as ethylene-responsive transcription factor CRF4-like and ABC transporter B family member 11-like, were significantly inhibited in response to EuPHB1 overexpression. Its overexpression disturbs the original signal transduction pathway, thus causing the corresponding phenotypic changes in transgenic tobacco. Indeed, such overexpression caused fading of palisade tissue and an increase in the number of certain mesophyll cells. It also increased adenosine triphosphate (ATP) synthase activity, mitochondrial membrane potential, ATP content, and reactive oxygen species (ROS) levels in cells. Our results suggest that EuPHB1 expression promotes cellular energy metabolism by accelerating the oxidative phosphorylation of the mitochondrial respiratory chain. Elevated levels of EuPHB1 in the mitochondria, which helps supply the extra energy required to support rapid rates of cell division.

Does atp8 exist in the mitochondrial genome of Proseriata (Metazoa: Platyhelminthes)?

The adenosine triphosphate (ATP) synthase F0 subunit 8 gene (atp8) had been believed to be absent in mitochondrial genomes of platyhelminths until the late 2010s, since when multiple lines of emergent evidence have suggested that this gene is actually present, albeit in highly derived forms, throughout the entire phylum except for the parasitic Neodermata (tapeworms, flukes, and their kin). Of about 11 non-parasitic (turbellarian) major platyhelminth subtaxa, the existence of atp8 has hitherto been documented in five (Catenulida, Macrostomorpha, Polycladida, Rhabdocoela, and Tricladida), while it remains open in the remaining six (Prorhynchida, Gnosonesmida, Proseriata, Fecampiida, Prolecithophora, and Bothrioplanida). Here we report the mitochondrial genome sequence of an undetermined marine interstitial turbellarian species in the genus NematoplanaMeixner, 1938 as the first representative of Proseriata. This circular genome comprises 16,106 bp (but potentially 18,812–19,277 bp when unresolved, non-coding tandem repeats are considered) and includes 38 genes, viz. 23 transfer RNA genes, 13 protein-coding genes (including the putative atp8), and two ribosomal RNA genes. The putative atp8 in Nematoplana sp. was not annotated by a standard automated procedure but was detected by manual inspection. If it encodes a translated product, it consists of 156 bp, with the potential 52-amino-acid-residue product beginning with MPHV, instead of the metazoan-canonical MPQL, and containing a single putative transmembrane region expanding from the 7th to the 29th amino-acid positions. While our finding seemingly strengthens the hypothesis that atp8 is in the ground pattern of flatworm mitochondrial genomes, whether the putative atp8 in flatworms is actually transcribed and translated to form a functional ATP synthetase F0 subunit should be tested in future studies.

Resilient and sustainable production of peanut (Arachis hypogaea) in phosphorus-limited environment by using exogenous gamma-aminobutyric acid to sustain photosynthesis

Globally, many low to medium yielding peanut fields have the potential for further yield improvement. Low phosphorus (P) limitation is one of the significant factors curtailing Arachis hypogaea productivity in many regions. In order to demonstrate the effects of gamma-aminobutyric acid (GABA) on peanuts growing under P deficiency, we used a pot-based experiment to examine the effects of exogenous GABA on alleviating P deficiency-induced physiological changes and growth inhibition in peanuts. The key physiological parameters examined were foliar gas exchange, photochemical efficiency, proton motive force, reactive oxygen species (ROS), and adenosine triphosphate (ATP) synthase activity of peanuts under cultivation with low P (LP, 0.5 mM P) and control conditions. During low P, the cyclic electron flow (CEF) maintained the high proton gradient (∆pH) induced by low ATP synthetic activity. Applying GABA during low P conditions stimulated CEF and reduced the concomitant ROS generation and thereby protecting the foliar photosystem II (PSII) from photoinhibition. Specifically, GABA enhanced the rate of electronic transmission of PSII (ETRII) by pausing the photoprotection mechanisms including non-photochemical quenching (NPQ) and ∆pH regulation. Thus, GABA was shown to be effective in restoring peanut growth when encountering P deficiency. Exogenous GABA alleviated two symptoms (increased root-shoot ratio and photoinhibition) of P-deficient peanuts. This is possibly the first report of using exogenous GABA to restore photosynthesis and growth under low P availability. Therefore, foliar applications of GABA could be a simple, safe and effective approach to overcome low yield imposed by limited P resources (low P in soils or P-fertilizers are unavailable) for sustainable peanut cultivation and especially in low to medium yielding fields.

Stem cell-derived exosome patch with coronary artery bypass graft restores cardiac function in chronically ischemic porcine myocardium

ObjectiveThis study aimed to investigate whether or not the application of a stem cell-derived exosome-laden collagen patch (EXP) during coronary artery bypass grafting (CABG) can recover cardiac function by modulating mitochondrial bioenergetics and myocardial inflammation in hibernating myocardium (HIB), which is defined as myocardium with reduced blood flow and function that retains viability and variable contractile reserve. MethodsIn vitro methods involved exposing H9C2 cardiomyocytes to hypoxia followed by normoxic coculture with porcine mesenchymal stem cells. Mitochondrial respiration was measured using Seahorse assay. GW4869, an exosomal release antagonist, was used to determine the effect of mesenchymal stem cells-derived exosomal signaling on cardiomyocyte recovery. Total exosomal RNA was isolated and differential micro RNA expression determined by sequencing. In vivo studies comprised 48 Yorkshire-Landrace juvenile swine (6 normal controls, 17 HIB, 19 CABG, and 6 CABG + EXP), which were compared for physiologic and metabolic changes. HIB was created by placing a constrictor on the proximal left anterior descending artery, causing significant stenosis but preserved viability by 12 weeks. CABG was performed with or without mesenchymal stem cells-derived EXP application and animals recovered for 4 weeks. Before terminal procedure, cardiac magnetic resonance imaging at rest, and with low-dose dobutamine, assessed diastolic relaxation, systolic function, graft patency, and myocardial viability. Tissue studies of inflammation, fibrosis, and mitochondrial morphology were performed posttermination. ResultsIn vitro data demonstrated improved cardiomyocyte mitochondrial respiration upon coculture with MSCs that was blunted when adding the exosomal antagonist GW4869. RNA sequencing identified 8 differentially expressed micro RNAs in normoxia vs hypoxia-induced exosomes that may modulate the expression of key mitochondrial (peroxisome proliferator-activator receptor gamma coactivator 1-alpha and adenosine triphosphate synthase) and inflammatory mediators (nuclear factor kappa-light-chain enhancer of activated B cells, interferon gamma, and interleukin 1β). In vivo animal magnetic resonance imaging studies demonstrated regional systolic function and diastolic relaxation to be improved with CABG + EXP compared with HIB (P = .02 and P = .02, respectively). Histologic analysis showed increased interstitial fibrosis and inflammation in HIB compared with CABG + EXP. Electron microscopy demonstrated increased mitochondrial area, perimeter, and aspect ratio in CABG + EXP compared with HIB or CABG alone (P < .0001). ConclusionsExosomes recovered cardiomyocyte mitochondrial respiration and reduced myocardial inflammation through paracrine signaling, resulting in improved cardiac function.

The mitochondrially targeted peptide elamipretide (SS-31) improves ADP sensitivity in aged mitochondria by increasing uptake through the adenine nucleotide translocator (ANT)

Aging muscle experiences functional decline in part mediated by impaired mitochondrial ADP sensitivity. Elamipretide (ELAM) rapidly improves physiological and mitochondrial function in aging and binds directly to the mitochondrial ADP transporter ANT. We hypothesized that ELAM improves ADP sensitivity in aging leading to rescued physiological function. We measured the response to ADP stimulation in young and old muscle mitochondria with ELAM treatment, in vivo heart and muscle function, and compared protein abundance, phosphorylation, and S-glutathionylation of ADP/ATP pathway proteins. ELAM treatment increased ADP sensitivity in old muscle mitochondria by increasing uptake of ADP through the ANT and rescued muscle force and heart systolic function. Protein abundance in the ADP/ATP transport and synthesis pathway was unchanged, but ELAM treatment decreased protein s-glutathionylation incuding of ANT. Mitochondrial ADP sensitivity is rapidly modifiable. This research supports the hypothesis that ELAM improves ANT function in aging and links mitochondrial ADP sensitivity to physiological function.Graphical abstractELAM binds directly to ANT and ATP synthase and ELAM treatment improves ADP sensitivity, increases ATP production, and improves physiological function in old muscles. ADP (adenosine diphosphate), ATP (adenosine triphosphate), VDAC (voltage-dependent anion channel), ANT (adenine nucleotide translocator), H+ (proton), ROS (reactive oxygen species), NADH (nicotinamide adenine dinucleotide), FADH2 (flavin adenine dinucleotide), O2 (oxygen), ELAM (elamipretide), –SH (free thiol), –SSG (glutathionylated protein)

MTA1, a Novel ATP Synthase Complex Modulator, Enhances Colon Cancer Liver Metastasis by Driving Mitochondrial Metabolism Reprogramming.

Liver metastasis is the most fatal event of colon cancer patients. Warburg effect has been long challenged by the fact of upregulated oxidative phosphorylation (OXPHOS), while its mechanism remains unclear. Here, metastasis-associated antigen 1 (MTA1) is identified as a newly identified adenosine triphosphate (ATP) synthase modulator by interacting with ATP synthase F1 subunit alpha (ATP5A), facilitates colon cancer liver metastasis by driving mitochondrial bioenergetic metabolism reprogramming, enhancing OXPHOS; therefore, modulating ATP synthase activity and downstream mTOR pathways. High-throughput screening of an anticancer drug shows MTA1 knockout increases the sensitivity of colon cancer to mitochondrial bioenergetic metabolism-targeted drugs and mTOR inhibitors. Inhibiting ATP5A enhances the sensitivity of liver-metastasized colon cancer to sirolimus in an MTA1-dependent manner. The therapeutic effects are verified in xenograft models and clinical cases. This research identifies a new modulator of mitochondrial bioenergetic reprogramming in cancer metastasis and reveals a new mechanism on upregulating mitochondrial OXPHOS as the reversal of Warburg effect in cancer metastasis is orchestrated.

A tale of two inhibitors: diarylquinolines and squaramides.

The diarylquinoline bedaquiline (BDQ) is an FDA-approved drug for the treatment of multidrug-resistant tuberculosis that targets the mycobacterial adenosine triphosphate (ATP) synthase, a key enzyme in cellular respiration. In a recent study, Courbon etal (2023) examine the interaction between Mycobacterium smegmatis ATP synthase with the second generation diarylquinoline TBAJ-876 and the squaramide inhibitor SQ31f, showing that both drugs prevent the rotatory motions needed for enzymatic function.