A5062359225- Force Microscopy Techniques and Applications
- Mechanical and Optical Resonators
- Advanced Electron Microscopy Techniques and Applications
- Lipid Membrane Structure and Behavior
- Molecular Junctions and Nanostructures
- Integrated Circuits and Semiconductor Failure Analysis
- Photoreceptor and optogenetics research
- Near-Field Optical Microscopy
- Advanced Fluorescence Microscopy Techniques
- Glycosylation and Glycoproteins Research
- Polymer Surface Interaction Studies
- Surface and Thin Film Phenomena
- Cellular transport and secretion
- Nanofabrication and Lithography Techniques
- Hydrogels: synthesis, properties, applications
- Monoclonal and Polyclonal Antibodies Research
- ATP Synthase and ATPases Research
- RNA and protein synthesis mechanisms
- Electrochemical Analysis and Applications
- Microtubule and mitosis dynamics
- Photosynthetic Processes and Mechanisms
- Cellular Mechanics and Interactions
- Supramolecular Self-Assembly in Materials
- Enzyme Structure and Function
- Advanced biosensing and bioanalysis techniques
Nagoya University
2017-2025
National Institutes of Natural Sciences
2019-2025
The Exploratory Research Center on Life and Living Systems
2018-2025
Kanazawa University
2010-2024
Institute for Molecular Science
2019-2024
Living Systems (United States)
2018-2024
Rigaku (Japan)
2023
Creative Research
2023
Graduate School USA
2019-2021
Aichi University
2018-2020
A deeper mechanistic understanding of the saccharification cellulosic biomass could enhance efficiency biofuels development. We report here real-time visualization crystalline cellulose degradation by individual cellulase enzymes through use an advanced version high-speed atomic force microscopy. Trichoderma reesei cellobiohydrolase I (TrCel7A) molecules were observed to slide unidirectionally along surface but at one point exhibited collective halting analogous a traffic jam. Changing...
F(1) is an adenosine triphosphate (ATP)-driven motor in which three torque-generating β subunits the α(3)β(3) stator ring sequentially undergo conformational changes upon ATP hydrolysis to rotate central shaft γ unidirectionally. Although extensive experimental and theoretical work has been done, structural basis of cooperative torque generation realize unidirectional rotation remains elusive. We used high-speed atomic force microscopy show that rotorless still "rotates"; isolated ring,...
The CRISPR-associated endonuclease Cas9 binds to a guide RNA and cleaves double-stranded DNA with sequence complementary the guide. Cas9-RNA system has been harnessed for numerous applications, such as genome editing. Here we use high-speed atomic force microscopy (HS-AFM) visualize real-space real-time dynamics of CRISPR-Cas9 in action. HS-AFM movies indicate that, whereas apo-Cas9 adopts unexpected flexible conformations, forms stable bilobed structure interrogates target sites on by...
Protein translocation across cellular membranes is an essential and nano-scale dynamic process. In the bacterial cytoplasmic membrane, core proteins in this process are a membrane protein complex, SecYEG, corresponding to eukaryotic Sec61 protein, SecA ATPase. Despite more than three decades of extensive research on Sec proteins, from genetic experiments cutting-edge single-molecule analyses, no study has visually demonstrated translocation. Here, we visualize translocation, via one unit...
Background and Objective: A major factor in the pathogenesis of periodontal disease, which is one biofilm infectious diseases, thought to be lipopolysaccharide (LPS), owing its ability cause inflammation promote tissue destruction. Moreover, elimination pathogens their component LPSs essential for successful treatment disease. Lipopolysaccharide tolerance a mechanism that prevents excessive prolonged responses monocytes macrophages LPS. Since persistence necessary inflammatory cytokine...
Abstract Light-driven outward H + pumps are widely distributed in nature, converting sunlight energy into proton motive force. Here we report the characterization of an oppositely directed pump with a similar architecture to pumps. A deep-ocean marine bacterium, Parvularcula oceani , contains three rhodopsins, one which functions as light-driven inward when expressed Escherichia coli and mouse neural cells. Detailed mechanistic analyses purified proteins reveal that small differences...
Abstract Motile cilia are microtubule-based organelles that play important roles in most eukaryotes. Although axonemal microtubules sufficiently stable to withstand their beating motion, it remains unknown how they stabilized while serving as tracks for dyneins. To address this question, we have identified two uncharacterized proteins, FAP45 and FAP52, microtubule inner proteins (MIPs) Chlamydomonas . These conserved among eukaryotes with motile cilia. Using cryo-electron tomography...
Visualization of morphological dynamics live cells with nanometer resolution under physiological conditions is highly desired, but challenging. It has been demonstrated that high-speed atomic force microscopy a powerful technique for visualizing biomolecules conditions. However, application imaging larger objects such as mammalian complicated because the collision between cantilever and samples. Here, we demonstrate attaching an extremely long (~3 μm) thin (~5 nm) tip by amorphous carbon to...
Oligomeric assembly is a common feature of membrane proteins and often relevant to their physiological functions. Determining the stoichiometry oligomeric state in lipid bilayer generally challenging because large size, complexity, structural alterations under experimental conditions. Here, we use high-speed atomic force microscopy (HS-AFM) directly observe states various microbial rhodopsins found within eubacteria archaea. HS-AFM images show that eubacterial predominantly exist as pentamer...
Schizorhodopsins, recently discovered rhodopsins in Asgard archaea, function as light-driven inward H + pumps.
The abuse of antibiotics has led to the emergence multidrug-resistant microbial pathogens, presenting a pressing challenge in global healthcare. Membrane-disrupting antimicrobial peptides (AMPs) combat so-called superbugs via mechanisms different than conventional and have good application prospects medicine, agriculture, food industry. However, mechanism-of-action AMPs not been fully characterized at cellular level due lack high-resolution imaging technologies that can capture...
Abstract The collision sensor Hel2 specifically recognizes colliding ribosomes and ubiquitinates the ribosomal protein uS10, leading to noncanonical subunit dissociation by ribosome-associated quality control trigger (RQT) complex. Although uS10 ubiquitination is essential for rescuing stalled ribosomes, its function recognition steps are not fully understood. Here, we show that RQT complex components Cue3 Rqt4 interact with K63-linked ubiquitin chain accelerate recruitment of ubiquitinated...
Combining a carbon nanotube probe and highly sensitive dynamic measurement scheme has enabled us to use atomic force microscopy measure oscillatory forces in water on approaching surface that been laterally characterized nanometer scale. This opens up the possibility of investigating layers under variety experimental conditions as function precise lateral position any including biological membranes macromolecules. Among many varied roles are effects biomolecular adhesion, colloid dispersion,...
Use of the atomic force microscope (AFM) in quantitative measurements inherently requires a theoretical framework enabling conversion observed deflection properties cantilever to an interaction force. In this paper, foundations using frequency modulation microscopy (FM-AFM) are examined and rigorously elucidated, with consideration being given both 'conservative' 'dissipative' interactions. This includes detailed discussion underlying assumptions involved such measurements, presentation...
We experimentally investigated a force interaction between Si tip and the Si(111)7\ifmmode\times\else\texttimes\fi{}7 reconstructed surface using noncontact-mode atomic-force microscope (AFM) in ultrahigh vacuum. Two types of gradient curves with without discontinuity were found. Furthermore, correlation noncontact AFM image contrasts has been clarified. The contrast was very weak case curve discontinuity, while it clearly enhanced inequivalent adatoms obtained discontinuity. can be...
Intrinsically disordered (ID) regions of proteins are recognized to be involved in biological processes such as transcription, translation, and cellular signal transduction. Despite the important roles ID regions, effective methods observe these thin flexible structures directly were not available. Herein, we use high-speed atomic force microscopy (AFM) heterodimeric FACT (facilitates chromatin transcription) protein, which is predicted have large each subunit. Successive AFM images on a...