Glucose Phosphorylation Research Articles

BMP4 aggravates mitochondrial dysfunction of HRMECs

Mitochondria are important places for the oxidative phosphorylation of glucose and the maintenance of cell oxidation and antioxidant stability. However, mitochondrial dysfunction causes cell dysfunction. Meanwhile, retinal vascular endothelial cell dysfunction may cause vascular inflammation, hemorrhage, angiogenesis, and other manifestations. Our previous studies have shown that Bone morphogenetic protein 4 (BMP4) is an important target for the treatment of retinal neovascularization, but the mechanism remains unclear. Therefore, our study aims to observe the effects of BMP4 on vascular endothelial cells and hopes to provide a new target for diabetic retinopathy. 4-Hydroxynonenal (4HNE), a kind of lipid peroxide, was used to induce the oxidative stress model. Human retinal microvascular endothelial cells (HRMECs) were randomly divided into control, 4HNE, negative control, and siBMP4 groups. Si-BMP4 significantly reduced leukocyte adhesion and 4HNE-induced high ROS level and restored the mitochondrial membrane potential and OCR. This indicates that BMP4 plays an important role in inducing leukocyte adhesion, oxidative stress, and mitochondrial dysfunction. The relationship between BMP4 and retinal vascular endothelial cell dysfunction is preliminarily confirmed by this study. Mitochondrial dysfunction and oxidative stress may be involved in BMP4-mediated retinal vascular endothelial cell dysfunction.

Exploring the prognostic value of HK3 and its association with immune infiltration in glioblastoma multiforme.

Background: Hexokinase 3 (HK3) is one of the key enzymes involved in glucose phosphorylation (the first step in most glucose metabolic pathways). Many studies have demonstrated the vital role of dysregulation of HK3 in several tumors. However, there is a need for in-depth characterization of the role of HK3 in glioblastoma multiforme (GBM). Methods: All data were sourced from The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA). Kaplan-Meier analysis and univariate regression were applied for survival analysis. Gene set enrichment analysis (GSEA) was used for enrichment analysis. Tumor Immune Single Cell Hub (TISCH) database was applied for single-cell analysis. Tumor Immune Dysfunction and Exclusion (TIDE) analysis was applied to evaluate the immune response. Results: HK3 expression was upregulated in GBM and correlated with poor prognosis. The high HK3 expression group was primarily enriched in adaptive immune response, chemokine signaling pathway, and cytokine-cytokine receptor interaction. The high HK3 expression group showed significantly greater enrichment of the majority of immune cells and immune-related pathways. HK3 showed significant correlation with most immune cells, especially macrophages (p < .001, R = .81). TISCH analysis showed that HK3 was predominantly expressed in macrophages in most cancers. HK3 showed significant correlation with most immune-related genes, such as PD-1 (p < .001, R = .41), PDL-1 (p < .001, R = .27), and CTLA-4 (p < .001, R = .29). TIDE analysis revealed that the low HK3 expression group has a lower TIDE score and may benefit from immunotherapy. Drug sensitivity analysis showed that patients with high HK3 expression frequently showed drug resistance. Conclusion: HK3 was associated with poor prognosis and may serve as a biomarker of macrophages in GBM. HK3 was also associated with immune response and drug resistance. Our findings may provide novel insights for GBM immunotherapy.

Role of phosphate dysmetabolism in the pathogenesis of chronic fatigue syndrome

Oxidative phosphorylation is a fundamental process of brain energy metabolism. It provides nearly all of the ATP used then in multiple phosphorylation cycles, including glucose phosphorylation. Moderate (40%) efficiency of the glucose ATP cycle requires a continuous extracellular supply of phosphates which deficiency is manifested as chronic fatigue

ANALYTIC SOLUTION OF ONE-DIMENSIONAL FRACTIONAL GLYCOLYSIS MODEL

Glycolysis, which occurs in the cytoplasm of both prokaryotic and eukaryotic cells, is regarded to be the primary step employed in the breakdown of glucose to extract energy. As it is utilized by all living things on the planet, it was likely one of the first metabolic routes to emerge. It is a cytoplasmic mechanism that converts glucose into carbon molecules while also producing energy. The enzyme hexokinase aids in the phosphorylation of glucose. Hexokinase is inhibited by this mechanism, which generates glucose-6-P from adenosine triphosphate (ATP). This paper’s primary goal is to quantitatively examine the general reaction–diffusion Glycolysis system. Since the Glycolysis model shows a positive result as the unknown variables represent chemical substance concentration, therefore, to evaluate the behavior of the model for the non-integral order derivative of both independent variables, we expanded the concept of the conventional order Glycolysis model to the fractional Glycolysis model. The nonlinearity of the model is decomposed through an Adomian polynomial for evaluation. More precisely, we used the iterative Laplace Adomian decomposition method (LADM) to determine the numerical solution for the underlying model. The model’s necessary analytical/numerical solution was found by adding the first few iterations. Finally, we have presented numerical examples and graphical representations to explain the dynamics of the considered model to ensure the scheme’s validity.

ИССЛЕДОВАНИЕ IN SILICO АФФИННОСТИ РИВАРОКСАБАНА К ГЛЮКОКИНАЗЕ

Type 2 diabetes mellitus is an urgent problem of health care. Comparison of the structure of the Rivaroxaban with the structure of a number of experimental drugs, which are Glucokinase activators, revealed the perspectives of its evalua-tion in the drug design for the treatment of type 2 diabetes. In in silico experi-ments, the interaction of Rivaroxaban with glucokinase, which is involved in glu-cose phosphorylation in liver and pancreas cells, has been studied. Using molecu-lar docking methods, complexes of Rivaroxaban with a protein in the allosteric center responsible for the action of glucokinase activators, were generated: the minimum binding energy was found 9.41 kcal/mol. The results obtained in the experiment in silico open the perspectives of design and development of the new effective antidiabetic drugs as well as investigation of the concomitant action of the anticoagulant Rivaroxaban.

A novel pathogenic variant in the glucokinase gene found in two Japanese siblings with maturity-onset diabetes of the young 2.

Glucokinase is a glycolytic enzyme that catalyzes the phosphorylation of glucose to glucose-6-phospate in the first step of the glycolytic pathway. It also regulates the threshold for insulin secretion from pancreatic beta cells by catalyzing the phosphorylation of glucose and plays an important role as a glucose sensor. Pathogenic variants in the glucokinase gene (GCK) cause non-progressive but persistent mild fasting hyperglycemia, also recognized as maturity-onset diabetes of the young 2 (MODY2). This report presents the case of two Japanese siblings with MODY2, who were initially diagnosed with impaired glucose intolerance at 20 and 17 years of age, and later developed diabetes mellitus. They had no history of obesity, were negative for islet-related autoantibodies and their serum C-peptide level were within the normal range. Diabetic complications were not observed. Next-generation sequencing revealed a novel heterozygous variant in GCK (NM_000162.5: c.1088A>G, p.Asp363Gly) in both siblings. This variant has not been reported previously. In silico functional analyses, using SIFT and MutationTaster, suggested that the variant was damaging. To confirm the functional impact of the mutated GCK, the HiBiT-tagged p.Asp363Gly variant and the wild-type GCK were transiently expressed in HEK293T cells. The cells expressing the variant GCK exhibited 79% less bioluminescence, compared to those expressing the wild-type GCK, suggesting that the pathophysiology of the variant was a result of haploinsufficiency.

Herpes Simplex Type 1 UL43 Multiple Membrane-Spanning Protein Increases Energy Metabolism in Host Cells through Interacting with ARL2.

Non-essential proteins for viral replication affect host cell metabolism, while the function of the UL43 protein of herpes simplex virus 1 (HSV-1) is not clear. Herein, we performed a comprehensive microarray analysis of HUVEC cells infected with HSV-1 and its UL43-deficient mutant and found significant variation in genes associated with cellular energy metabolic pathways. The localization of UL43 protein in host cells and how it affects cellular energy metabolism pathways were further investigated. Internalization analysis showed that the UL43 protein could be endocytosis-mediated by YPLF motif (aa144-147) and localized to mitochondria. At the same time, more ATP was produced by coupling with mitochondrial small G protein ARF-like 2 (ARL2) GTPase, which triggered the phosphorylation of ANT1 (SLC25A4) to affect the opening degree of mitochondrial permeability transition pore (mPTP), and significantly promoted the aerobic oxidation and oxidative phosphorylation of glucose. Our study shows that UL43 mediates the improvement of host cell metabolism after HSV-1 infection. Additionally, UL43 protein could be a valuable ATP-stimulating factor for mammalian cells.

Aberrant Energy Metabolism in Alzheimer's Disease.

To maintain energy supply to the brain, a direct energy source called adenosine triphosphate (ATP) is produced by oxidative phosphorylation and aerobic glycolysis of glucose in the mitochondria and cytoplasm. Brain glucose metabolism is reduced in many neurodegenerative diseases, including Alzheimer's disease (AD), where it appears presymptomatically in a progressive and region-specific manner. Following dysregulation of energy metabolism in AD, many cellular repair/regenerative processes are activated to conserve the energy required for cell viability. Glucose metabolism plays an important role in the pathology of AD and is closely associated with the tricarboxylic acid cycle, type 2 diabetes mellitus, and insulin resistance. The glucose intake in neurons is from endothelial cells, astrocytes, and microglia. Damage to neurocentric glucose also damages the energy transport systems in AD. Gut microbiota is necessary to modulate bidirectional communication between the gastrointestinal tract and brain. Gut microbiota may influence the process of AD by regulating the immune system and maintaining the integrity of the intestinal barrier. Furthermore, some therapeutic strategies have shown promising therapeutic effects in the treatment of AD at different stages, including the use of antidiabetic drugs, rescuing mitochondrial dysfunction, and epigenetic and dietary intervention. This review discusses the underlying mechanisms of alterations in energy metabolism in AD and provides potential therapeutic strategies in the treatment of AD.

Energetic insufficiency of liver tissue and additional energy in digital value. (The hypothesis)

Despite the deep understanding of the importance of energy supply in the development of the vast majority of processes and phenomena in organism, there are practically no conceptual researches of energy requirements in severe diseases, traumatic injuries, stressful effects and related treatment in the medical literature.The proposed hypothesis is based on the analysis of literature data (PubMed keywords: energy deficit, stressor, metabolic stress, ATP, gluconeogenesis, oxidative phosphorylation, mitochondria, insulin secretion), reconsideration of the results of our experiments dedicated to the energetic statement of liver tissue in obstructive jaundice (OJ), ischemia and massive resection, summarization of 60 years of experience in clinical, surgical and scientific activities, which made it possible to make a number of assumptions that need further clinical and experimental verification.Various pathogens (stressors) cause the additional energy production in the body, which is the energy basis of metabolic responses that ensure the adaptation of the body's vital functions and the elimination of the pathogen by activation of innate immunity, systemic inflammatory reaction, activation of the sympathetic nervous system, etc.Additional energy is the integral strength of the response to the pathogen, that takes into consideration with the strength of the stressor and the individual strength of the body's response, which can be different for the same strength of the stressor. In fact, when stress develops, it determines its strength in digital form, i.e. in real view.The concept of this hypothesis comes from the fact that stress, which main task is to provide the energy of organism, appears when there is a certain level of energy deficiency in the body.Such level rarely appears immediately after the action of the stressor. At the beginning, pre-stress adaptive reactions usually occur, which use the energy reserve in cells in the form of ATP and glycogen, and are also the result of energy redistribution: a decrease in it in insulin-dependent tissues and an increase in insulin-independent ones, which include vital organs.This made it possible to divide metabolic responses into two groups: pre-stress and stress, and to distinguish two periods: “pre-stress” and “metabolic stress”. Pre-stress reactions, in our opinion, are also aimed at preventing the development of metabolic stress, which generates energy through proteolysis and lipolysis of body tissues. Metabolic stress develops when pre-stress reactions cannot satisfy the needs of the body and a certain, expressed in numerical value, energy deficit occurs. In a certain extent the metabolic situation in the body reflects by the liver, which is a metabolic organ that performs many reactions both during normal and stressful metabolism, generates ATP energy, and takes into account the metabolic state of other organs. The level of energy deficiency of the liver tissue can be an indicator that causes the formation of metabolic stress and evaluates in a numerical value not only the energy position of the body, but also the severity of its general condition, promising opportunities, prognosis and priority treatment, which should be aimed at a comprehensive replenishment of the energy deficit.This is especially important to keep in mind at this time with severe forms of COVID-19 and low blood oxygen saturation. Under any stressful influences, the doctor must solve two problems: to deal with a specific stressor and to provide energy for this struggle and the vital activity of the patient. Functional insufficiency of the liver in its diffuse diseases can lead to impaired gluconeogenesis or oxidative phosphorylation of glucose and the formation of “unsuccessful” or “incomplete” stresses.

Reduction in Insulin Mediated ERK Phosphorylation by Palmitate in Liver Cells Is Independent of Fatty Acid Induced ER Stress.

Saturated free fatty acids (FFAs) such as palmitate in the circulation are known to cause endoplasmic reticulum (ER) stress and insulin resistance in peripheral tissues. In addition to protein kinase B (AKT) signaling, extracellular signal-regulated kinase (ERK) has been implicated in the development of insulin resistance. However, there are conflicting data regarding role of ERK signaling in ER stress-induced insulin resistance. In this study, we investigated the effects of ER stress on insulin resistance and ERK phosphorylation in Huh-7 cells and evaluated how oleate prevents palmitate-mediated ER stress. Treatment with insulin resulted in an increase of 38–45% in the uptake of glucose in control cells compared to non-insulin-treated control cells, along with an increase in the phosphorylation of AKT and ERK. We found that treatment with palmitate increased the expression of ER stress genes, including the splicing of X box binding protein 1 (XBP1) mRNA. At the same time, we observed a decrease in insulin-mediated uptake of glucose and ERK phosphorylation in Huh-7 cells, without any change in AKT phosphorylation. Supplementation of oleate along with palmitate mitigated the palmitate-induced ER stress but did not affect insulin-mediated glucose uptake or ERK phosphorylation. The findings of this study suggest that palmitate reduces insulin-mediated ERK phosphorylation in liver cells and this effect is independent of fatty-acid-induced ER stress.

The effect of inter-varietal variation in sugar hydrolysis and transport on sugar content and photosynthesis in Vitis vinifera L. leaves

Sugar synthesis from photosynthesis and its utilization through sugar metabolism jointly determine leaf sugar content, and in contrast, excess sugar represses leaf photosynthesis. Although plant photosynthesis is affected by leaf sugar metabolism, the relationship between sugar metabolism and photosynthetic capacity of different grape genotypes remains unclear. In this study, two grape (Vitis vinifera L.) genotypes ‘Riesling’ (RI, high sugar content in leaf) and ‘Petit Manseng’ (PM, low sugar content in leaf) were used to evaluate the relationship between sugar metabolism and photosynthesis. Sugar content, chlorophyll content, photosynthetic parameters, enzyme activity, and gene expression related to sucrose metabolism in leaves were measured, and the correlations between photosynthesis and sugar metabolism were assessed. The contents of sucrose and glucose were significantly higher in RI leaves than in PM leaves, while the fructose content pattern was reversed. Cell wall invertase activity for sucrose hydrolysis and the transcript levels of VvCWINV, VvHTs, VvTMT1, VvFKs, and VvHXK2 were also higher in RI leaves than in PM leaves, whereas that of VvHXK1 mediating glucose phosphorylation, was lower in RI leaves than in PM leaves. Net photosynthetic rate, stomatal conductance, transpiration rate, and chlorophyll content were lower in RI leaves than in PM leaves and negatively correlated with glucose content, and the transcript levels of VvCWINV, VvHTs, VvTMT1, and VvHXK2. In conclusion, this study indicates that leaf sugar metabolism and transport are related to photosynthesis in Vitis vinifera L., which provides a theoretical basis for improving grape photosynthesis.

OsHXK3 encodes a hexokinase-like protein that positively regulates grain size in rice.

We report the map-based cloning and functional characterization of SNG1, which encodes OsHXK3, a hexokinase-like protein that plays a pivotal role in controlling grain size in rice. Grain size is an important agronomic trait determining grain yield and appearance quality in rice. Here, we report the discovery of rice mutant short and narrow grain1 (sng1) with reduced grain length, width and weight. Map-based cloning revealed that the mutant phenotype was caused by loss of function of gene OsHXK3 that encodes a hexokinase-like (HKL) protein. OsHXK3 was associated with the mitochondria and was ubiquitously distributed in various organs, predominately in younger organs. Analysis of glucose (Glc) phosphorylation activities in young panicles and protoplasts showed that OsHXK3 was a non-catalytic hexokinase (HXK). Overexpression of OsHXK3 could not complement the Arabidopsis glucose insensitive2-1 (gin2-1) mutant, indicating that OsHXK3 lacked Glc signaling activity. Scanning electron microscopy analysis revealed that OsHXK3 affects grain size by promoting spikelet husk cell expansion. Knockout of other nine OsHXK genes except OsHXK3 individually did not change grain size, indicating that functions of OsHXKs have differentiated in rice. OsHXK3 influences gibberellin (GA) biosynthesis and homeostasis. Compared with wild type, OsGA3ox2 was significantly up-regulated and OsGA2ox1 was significantly down-regulated in young panicle of sng1, and concentrations of biologically active GAs were significantly decreased in young panicles of the mutants. The yield per plant of OsHXK3 overexpression lines (OE-4 and OE-35) was increased by 10.91% and 7.62%, respectively, compared to that of wild type. Our results provide evidence that an HXK lacking catalytic and sensory functions plays an important role in grain size and has the potential to increase yield in rice.

New Insights into Xanthophylls and Lipidomic Profile Changes Induced by Glucose Supplementation in the Marine Diatom Nitzschia laevis.

Nitzschia laevis is a candidate microorganism for bioactive compounds (fucoxanthin and eicosapentaenoic acid (EPA)) production. In this study, the impacts of glucose-induced trophic transition on biomass, photosynthesis, pigments, and lipid profiles were examined. The specific growth rate was increased under glucose addition, achieved at 0.47 day−1 (0.26 ± 0.01 day−1 for the group without glucose in medium). However, the photosynthetic parameters and pigments including chlorophylls, fucoxanthin, and diatoxanthin were reduced. The net yield of EPA doubled under glucose addition, reaching 20.36 ± 1.22 mg/L in 4 days. In addition, the alteration in detailed lipid molecular species was demonstrated with a focus on EPA-enriched lipids. The effects of 2-deoxyglucose (2DG) indicated that glucose phosphorylation was involved in glucose-induced regulation. These findings provide novel data for guiding the application of this diatom strain in the functional food industries.

New non‐canonical function of catalytically dead hexokinase 1 in ovarian cancer

Hexokinases (HKs) have been found amplified in primary cancer cells, with the amplification of HK1 reported commonly in ovarian cancer cells. The function of HKs extends beyond glucose phosphorylation, and HKs serve as regulatory proteins in tumorigenesis. We addressed the non‐canonical roles of HK1 in ovarian cancer cells by examining the effects of the deletions of HK1 and HK2 in TOV‐112D ovarian adenocarcinoma cells. We reverted these effects by re‐introducing wild‐type HK1 and HK2, and we compared the HK1 revertant with the knock‐in of catalytically‐dead HK1 p.D656A. We subjected these cells to a battery of metabolic and proliferation assays and targeted GCxGC‐MS metabolomics. When expressed in vitro, the p.D656A mutant did not manifest a measurable KM and exhibited Vmax lower by almost two orders of magnitude when compared to the wild‐type enzyme. Knock‐in of HK1 p.D656A into HK1 KO cells reverted the HK1 KO‐induced effects in a broad range of assays, including the alamarBlue cell viability assay under low glucose conditions, the NADPH/NADP and NAD/NADH ratios, and glutathione levels in metformin‐treated cells. In the presence of 0.4 mM glucose, the phenotype of HK1 p.D656A knock‐in cells resembled the phenotype of cells with deleted HK2 but not those with deleted HK1 with regards to the levels of TCA intermediates, aspartate and cysteine, and glutamate. Moreover, HK1 deletion increased the levels of branched amino acids, which was completely eliminated by the expression of catalytically‐dead HK1. The HK1 deletion (but not HK2 deletion) suppressed the growth of xenotransplanted ovarian cancer cells, and nearly abolished the tumor growth when the mice were fed the glucose‐free diet. In summary, we provided the evidence that HK1 is involved in the so far unknown glycolysis‐independent moonlighting function and influences metabolism even in the glucose‐free conditions.

Hexokinase 3 enhances myeloid cell survival via non-glycolytic functions

The family of hexokinases (HKs) catalyzes the first step of glycolysis, the ATP-dependent phosphorylation of glucose to glucose-6-phosphate. While HK1 and HK2 are ubiquitously expressed, the less well-studied HK3 is primarily expressed in hematopoietic cells and tissues and is highly upregulated during terminal differentiation of some acute myeloid leukemia (AML) cell line models. Here we show that expression of HK3 is predominantly originating from myeloid cells and that the upregulation of this glycolytic enzyme is not restricted to differentiation of leukemic cells but also occurs during ex vivo myeloid differentiation of healthy CD34+ hematopoietic stem and progenitor cells. Within the hematopoietic system, we show that HK3 is predominantly expressed in cells of myeloid origin. CRISPR/Cas9 mediated gene disruption revealed that loss of HK3 has no effect on glycolytic activity in AML cell lines while knocking out HK2 significantly reduced basal glycolysis and glycolytic capacity. Instead, loss of HK3 but not HK2 led to increased sensitivity to ATRA-induced cell death in AML cell lines. We found that HK3 knockout (HK3-null) AML cells showed an accumulation of reactive oxygen species (ROS) as well as DNA damage during ATRA-induced differentiation. RNA sequencing analysis confirmed pathway enrichment for programmed cell death, oxidative stress, and DNA damage response in HK3-null AML cells. These signatures were confirmed in ATAC sequencing, showing that loss of HK3 leads to changes in chromatin configuration and increases the accessibility of genes involved in apoptosis and stress response. Through isoform-specific pulldowns, we furthermore identified a direct interaction between HK3 and the proapoptotic BCL-2 family member BIM, which has previously been shown to shorten myeloid life span. Our findings provide evidence that HK3 is dispensable for glycolytic activity in AML cells while promoting cell survival, possibly through direct interaction with the BH3-only protein BIM during ATRA-induced neutrophil differentiation.

Altered Germinal-Center Metabolism in B Cells in Autoimmunity.

B lymphocytes play an important role in the pathophysiology of many autoimmune disorders by producing autoantibodies, secreting cytokines, and presenting antigens. B cells undergo extreme physiological changes as they develop and differentiate. Aberrant function in tolerogenic checkpoints and the metabolic state of B cells might be the contributing factors to the dysfunctionality of autoimmune B cells. Understanding B-cell metabolism in autoimmunity is important as it can give rise to new treatments. Recent investigations have revealed that alterations in metabolism occur in the activation of B cells. Several reports have suggested that germinal center (GC) B cells of individuals with systemic lupus erythematosus (SLE) have altered metabolic function. GCs are unique microenvironments in which the delicate and complex process of B-cell affinity maturation occurs through somatic hypermutation (SHM) and class switching recombination (CSR) and where Bcl6 tightly regulates B-cell differentiation into memory B-cells or plasma cells. GC B cells rely heavily on glucose, fatty acids, and oxidative phosphorylation (OXPHOS) for their energy requirements. However, the complicated association between GC B cells and their metabolism is still not clearly understood. Here, we review several studies of B-cell metabolism, highlighting the significant transformations that occur in GC progression, and suggest possible approaches that may be investigated to more precisely target aberrant B-cell metabolism in SLE.

A perylene diimide-based nanoring architecture for exogenous and endogenous ATP detection: biochemical assay for monitoring phosphorylation of glucose.

Positively charged amphiphiles hold great significance in supramolecular chemistry due to their good solubility, and physiochemical and molecular recognition properties. Herein, we report the synthesis, characterization and molecular recognition properties of the dicationic amphiphile based on perylene diimide-tyrosine alkyl amide amine (PDI 3). PDI 3 showed the formation of a nanoring architecture in the self-assembled aggregated state (90% H2O-DMSO mixture) as observed by SEM and TEM studies. The diameter of the nanoring is around 30-50 nm with a height varying from 1 to 2 nm. The self-assembled aggregates of PDI 3 are very sensitive towards nucleoside triphosphates. Upon addition of ATP, PDI 3 showed a decrease in the absorbance and emission intensity at 535 and 580 nm (due to the monomer state), respectively. The lowest detection limit for ATP is 10.8 nM (UV) and 3.06 nM (FI). Upon interaction of ATP with PDI 3, the nanoring morphology transformed into a spherical structure. These changes could be attributed to the formation of ionic self-assembled aggregates between dicationic PDI 3 and negatively charged ATP via electrostatic and H-bonding interactions. The complexation mechanism of PDI 3 and ATP was confirmed by optical, NMR, Job's plot, DLS, SEM and AFM studies. PDI 3 displays low cytotoxicity toward MG-63 cells and can be successfully used for the detection of exogenous and endogenous ATP. The resulting PDI 3 + ATP complex is successfully used as a 'turn-on' biochemical assay for monitoring phosphorylation of glucose.

Mitochondrial Metabolism in Myocardial Remodeling and Mechanical Unloading: Implications for Ischemic Heart Disease

Ischemic heart disease refers to myocardial degeneration, necrosis, and fibrosis caused by coronary artery disease. It can lead to severe left ventricular dysfunction (LVEF ≤ 35–40%) and is a major cause of heart failure (HF). In each contraction, myocardium is subjected to a variety of mechanical forces, such as stretch, afterload, and shear stress, and these mechanical stresses are clinically associated with myocardial remodeling and, eventually, cardiac outcomes. Mitochondria produce 90% of ATP in the heart and participate in metabolic pathways that regulate the balance of glucose and fatty acid oxidative phosphorylation. However, altered energetics and metabolic reprogramming are proved to aggravate HF development and progression by disturbing substrate utilization. This review briefly summarizes the current insights into the adaptations of cardiomyocytes to mechanical stimuli and underlying mechanisms in ischemic heart disease, with focusing on mitochondrial metabolism. We also discuss how mechanical circulatory support (MCS) alters myocardial energy metabolism and affects the detrimental metabolic adaptations of the dysfunctional myocardium.

MitomiRs: their roles in mitochondria and importance in cancer cell metabolism.

BackgroundMicroRNAs (miRNAs) are short non-coding RNAs that play important roles in almost all biological pathways. They regulate post-transcriptional gene expression by binding to the 3’untranslated region (3’UTR) of messenger RNAs (mRNAs). MitomiRs are miRNAs of nuclear or mitochondrial origin that are localized in mitochondria and have a crucial role in regulation of mitochondrial function and metabolism. In eukaryotes, mitochondria are the major sites of oxidative metabolism of sugars, lipids, amino acids, and other bio-macromolecules. They are also the main sites of adenosine triphosphate (ATP) production.ConclusionsIn the review, we discuss the role of mitomiRs in mitochondria and introduce currently well studied mitomiRs, their target genes and functions. We also discuss their role in cancer initiation and progression through the regulation of mRNA expression in mitochondria. MitomiRs directly target key molecules such as transporters or enzymes in cell metabolism and regulate several oncogenic signaling pathways. They also play an important role in the Warburg effect, which is vital for cancer cells to maintain their proliferative potential. In addition, we discuss how they indirectly upregulate hexokinase 2 (HK2), an enzyme involved in glucose phosphorylation, and thus may affect energy metabolism in breast cancer cells. In tumor tissues such as breast cancer and head and neck tumors, the expression of one of the mitomiRs (miR-210) correlates with hypoxia gene signatures, suggesting a direct link between mitomiR expression and hypoxia in cancer. The miR-17/92 cluster has been shown to act as a key factor in metabolic reprogramming of tumors by regulating glycolytic and mitochondrial metabolism. This cluster is deregulated in B-cell lymphomas, B-cell chronic lymphocytic leukemia, acute myeloid leukemia, and T-cell lymphomas, and is particularly overexpressed in several other cancers. Based on the current knowledge, we can conclude that there is a large number of miRNAs present in mitochondria, termed mitomiR, and that they are important regulators of mitochondrial function. Therefore, mitomiRs are important players in the metabolism of cancer cells, which need to be further investigated in order to develop a potential new therapies for cancer.

A mathematical model of enzymatic non-competitive inhibition by product and its applications

Enzymes are biological catalysts naturally present in living organisms, and they are capable of accelerating biochemical reactions in the metabolism process. Cells use many regulatory mechanisms to regulate the concentrations of cellular metabolites at physiological levels. Enzymatic inhibition is one of the key regulatory mechanisms naturally occurring in cellular metabolism, especially the enzymatic non-competitive inhibition by product. This inhibition process helps the cell regulate enzymatic activities. In this paper, we develop a novel mathematical model describing the enzymatic non-competitive inhibition by product. The model consists of a coupled system of nonlinear ordinary differential equations for the species of interest. Using nondimensionalization analysis, a formula for product formation rate for this mechanism is obtained in a transparent manner. Further analysis for this formula yields qualitative insights into the maximal reaction velocity and apparent Michaelis-Menten constant. Asymptotic solutions of the model are carefully given by using the homotopy perturbation analysis. A good agreement between the asymptotic solutions and numerical solutions are found. In addition, a Sobol global sensitivity analysis is implemented to help identify the key mechanisms of the enzyme activities. The results of this analysis show that the rate of product formation is relatively sensitive to the following factors: the catalytic rate of the enzyme, the rates of binding/unbinding of the product to/from the enzyme/enzyme complex. The numerical simulations provide insights into how variations in the model parameters affect the model output. Finally, an application of the model to the phosphorylation of glucose by mutant-hexokinase I enzyme is briefly discussed.