A5015613537- ATP Synthase and ATPases Research
- Mitochondrial Function and Pathology
- Cancer, Hypoxia, and Metabolism
- Enzyme Production and Characterization
- Adipose Tissue and Metabolism
- Microbial Metabolites in Food Biotechnology
- Photosynthetic Processes and Mechanisms
- Advanced Fluorescence Microscopy Techniques
- Neuroscience and Neuropharmacology Research
- Metabolism and Genetic Disorders
- Endoplasmic Reticulum Stress and Disease
- Adenosine and Purinergic Signaling
- Metalloenzymes and iron-sulfur proteins
- Metabolism, Diabetes, and Cancer
- Pancreatic function and diabetes
- Phagocytosis and Immune Regulation
- RNA and protein synthesis mechanisms
- Biochemical and Molecular Research
- Advanced Electron Microscopy Techniques and Applications
- Carbohydrate Chemistry and Synthesis
- Autophagy in Disease and Therapy
- Nanoplatforms for cancer theranostics
- Receptor Mechanisms and Signaling
- Enzyme Structure and Function
- Advanced MRI Techniques and Applications
Yamaguchi University
2024
Kyoto University
2015-2024
Leipzig University
2017
Japan Science and Technology Agency
2003-2013
The University of Tokyo
1990-2012
Osaka University
2006-2012
Kyoto Sangyo University
2012
Osaka Research Institute of Industrial Science and Technology
2007-2011
Imamura Hospital
2009
Osaka Prefectural Medical Center
2007
Adenosine 5'-triphosphate (ATP) is the major energy currency of cells and involved in many cellular processes. However, there no method for real-time monitoring ATP levels inside individual living cells. To visualize levels, we generated a series fluorescence resonance transfer (FRET)-based indicators that were composed epsilon subunit bacterial F(o)F(1)-ATP synthase sandwiched by cyan- yellow-fluorescent proteins. The indicators, named ATeams, had apparent dissociation constants ranging...
Recent advances in quantitative single-cell analysis revealed large diversity gene expression levels between individual cells, which could affect the physiology and/or fate of each cell. In contrast, for most metabolites, concentrations were only measureable as ensemble averages many cells. living adenosine triphosphate (ATP) is a critically important metabolite that powers intracellular reactions. Quantitative measurement absolute ATP concentration cells has not been achieved because lack...
V o 1 -ATPase is responsible for acidification of eukaryotic intracellular compartments and ATP synthesis Archaea some eubacteria. From the similarity to F -ATP synthase, has been assumed be a rotary motor, but date there are no experimental data support this. Here we visualized rotation single molecules -ATPase, catalytic subcomplex -ATPase. from Thermus thermophilus was immobilized onto glass surface, bead attached D or subunit through biotin-streptavidin linkage. In both cases observed...
Intracellular Ca2+ levels play a crucial role in the control of ATP synthesis. However, spatiotemporal correlation between and remains unclear due to inability visualize these factors within same individual cells. A Förster resonance energy transfer (FRET)-based fluorescent probe, named ATeam, was recently developed for imaging single living spectra cyan protein (CFP) yellow (YFP) used as FRET donor acceptor, respectively, significantly overlap with ultraviolet-excitable fura-2. In present...
For cell division, negatively charged chromatin, in which nucleosome fibers (10 nm fibers) are irregularly folded [1Eltsov M. Maclellan K.M. Maeshima K. Frangakis A.S. Dubochet J. Analysis of cryo-electron microscopy images does not support the existence 30-nm chromatin mitotic chromosomes situ.Proc. Natl. Acad. Sci. USA. 2008; 105: 19732-19737Crossref PubMed Scopus (282) Google Scholar, 2Nishino Y. Eltsov Joti Ito Takata H. Takahashi Hihara S. Imamoto N. Ishikawa T. Human consist...
DNA damage signaling is critical for the maintenance of genome integrity and cell fate decision. Poly(ADP-ribose) polymerase 1 (PARP1) a sensor rapidly activated in dose- complexity-dependent manner playing role initial chromatin organization repair pathway choice at sites. However, our understanding cell-wide consequence its activation damaged cells still limited. Using phasor approach to fluorescence lifetime imaging microscopy fluorescence-based biosensors combination with laser...
In several neurodegenerative diseases and myelin disorders, the degeneration profiles of myelinated axons are compatible with underlying energy deficits. However, it is presently impossible to measure selectively axonal ATP levels in electrically active nervous system. We combined transgenic expression an ATP-sensor neurons mice confocal FRET imaging electrophysiological recordings acutely isolated optic nerves. This allowed us monitor dynamic changes activity-dependent homeostasis at...
Adenosine 5′-triphosphate (ATP) is the major energy currency of all living organisms. Despite its important functions, spatiotemporal dynamics ATP levels inside multicellular organisms unclear. In this study, we modified genetically encoded Förster resonance transfer (FRET)-based biosensor ATeam to optimize affinity at low temperatures. This new biosensor, AT1.03NL, detected changes Drosophila S2 cells more sensitively than original did, 25 °C. By expressing AT1.03NL in melanogaster and...
Glucose is the mandatory fuel for brain, yet relative contribution of glucose and lactate neuronal energy metabolism unclear. We found that increased lactate, but not concentration, enhances spiking activity neurons cerebral cortex. Enhanced was dependent on ATP-sensitive potassium (K ATP ) channels formed with KCNJ11 ABCC8 subunits, which we show are functionally expressed in most neocortical types. also demonstrate ability cortical to take-up metabolize lactate. further reveal produced by...
Adenosine triphosphate (ATP) at millimolar levels has recently been implicated in the solubilization of cellular proteins. However, significance this high ATP level under physiological conditions and mechanisms that maintain remain unclear. We herein demonstrated AMP-activated protein kinase (AMPK) adenylate (ADK) cooperated to regardless glucose levels. Single-cell imaging ATP-reduced yeast mutants revealed these underwent stochastic transient depletion, which promoted cytotoxic aggregation...
Abstract Empagliflozin, a sodium-glucose co-transporter 2 inhibitor developed, has been shown to reduce cardiovascular events in patients with type diabetes and established disease. Several studies have suggested that empagliflozin improves the cardiac energy state which is partial cause of its potency. However, detailed mechanism remains unclear. To address this issue, we used mouse model enabled direct measurement cytosolic mitochondrial ATP levels. Empagliflozin treatment significantly...
The vacuole-type ATPases (V-ATPases) exist in various intracellular compartments of eukaryotic cells to regulate physiological processes by controlling the acidic environment. crystal structure subunit C Thermus thermophilus V-ATPase, homologous d V-ATPases, has been determined at 1.95-A resolution and located into holoenzyme complex obtained single particle analysis as suggested results cross-linking experiments. result shows that V-ATPase is substantially longer than related F-type ATPase,...
Fluorescent protein (FP)-based Forster resonance energy transfer (FRET) technology is useful for development of functional indicators to visualize second messenger molecules and activation signaling components in living cells. However, the design construction require careful optimization their structure at atomic level. Therefore, routine procedures constructing FRET-based currently include adjustment linker length between FPs sensor domain relative dipole orientation FP chromophore. Here we...
Cytosolic Ca(2+) signals are transferred into mitochondria over a huge concentration range. In our recent work we described uncoupling proteins 2 and 3 (UCP2/3) to be fundamental for mitochondrial uptake of high domains in mitochondria-ER junctions. On the other hand, leucine zipper EF hand-containing transmembrane protein 1 (Letm1) was identified as Ca(2+)/H(+) antiporter that achieved sequestration at small increases. Thus, contributions Letm1 UCP2/3 were compared endothelial cells....
Adenosine 5′-triphosphate (ATP) is the primary energy currency of all living organisms and participates in a variety cellular processes. Although ATP requirements during viral lifecycles have been examined number studies, method by which production can be monitored real-time, quantified individual cells subcellular compartments, lacking, thereby hindering studies aimed at elucidating precise mechanisms replication energized controlled. In this study, we investigated fluctuation distribution...
Significance We developed a sensitive method to assess the activity of oxidative phosphorylation in living cells using FRET-based ATP biosensor. then revealed that G0/G1 switch gene 2, protein rapidly induced by hypoxia, increases mitochondrial production interacting with F o 1 -ATP synthase and protects from critical energy crisis.
Abstract ATP levels may represent fundamental health conditions of cells. However, precise measurement intracellular in living cells is hindered by the lack suitable methodologies. Here, we developed a novel biosensor termed “BTeam”. BTeam comprises yellow fluorescent protein (YFP), binding domain ε subunit bacterial synthase, and an ATP-nonconsuming luciferase (NLuc). To attain emission, simply required NLuc substrate. showed elevated bioluminescence resonance energy transfer efficiency...
Vascular endothelial cells (ECs) sense and transduce hemodynamic shear stress into intracellular biochemical signals, Ca2+ signaling plays a critical role in this mechanotransduction, i.e., ECs release ATP the caveolae response to and, turn, released activates P2 purinoceptors, which results an influx of extracellular Ca2+. However, mechanism by evokes remains unclear. Here, we demonstrated that cellular mitochondria play process. Cultured human pulmonary artery were exposed controlled...
Abstract Differentiated podocytes, a type of renal glomerular cells, require substantial levels energy to maintain physiology. Mitochondria and glycolysis are two major producers ATP, but the precise roles each in podocytes remain unknown. This study evaluated mitochondria differentiated differentiating podocytes. located central part cell body while blocking had minor effects on shape migratory ability. In contrast, significantly reduced formation lamellipodia, cortical area these decreased...
Scientific Report7 March 2014Open Access Source Data Toll-like receptor 9 protects non-immune cells from stress by modulating mitochondrial ATP synthesis through the inhibition of SERCA2 Yasunori Shintani Corresponding Author William Harvey Research Institute, Barts and The London School Medicine Dentistry, Queen Mary University London, UK Department Medical Biochemistry, Osaka Graduate Medicine, Suita, Japan Search for more papers this author Hannes CA Drexler Bioanalytical Mass...
Highlights•We describe two types of ATP regulators: one limits consumption, while the other enhances production.•ATP regulators maintain levels and protect neuronal cells from ER stress eventual cell death.•ATP mitigate disease phenotypes in mouse models Parkinson's disease.ATP is essential human activities health. We hypothesize that maintaining may help to preserve vulnerable brain neurodegenerative diseases like (PD). For this purpose, we developed tested chemical compounds, for limiting...
Significance The mechanotransduction of shear stress in vascular endothelial cells is still not completely understood. We show a pathway signal transduction mediated by plasma membrane cholesterol-dependent mitochondrial oxidative phosphorylation. latest imaging technology using domain 4 mutant-derived cholesterol biosensors and fluorescence resonance energy transfer-based adenosine triphosphate (ATP) biosensor revealed that rapidly decreases levels the via both efflux internalization,...