A5034614522- Force Microscopy Techniques and Applications
- SARS-CoV-2 and COVID-19 Research
- Lipid Membrane Structure and Behavior
- Nuclear Structure and Function
- RNA Research and Splicing
- Extracellular vesicles in disease
- Nanopore and Nanochannel Transport Studies
- RNA and protein synthesis mechanisms
- Bacterial Infections and Vaccines
- Genomics and Chromatin Dynamics
- Bacteriophages and microbial interactions
- Advanced Electron Microscopy Techniques and Applications
Kanazawa University
2021-2025
Life Science Institute
2023
Abstract SARS‐CoV‐2 spike protein (S) binds to human angiotensin‐converting enzyme 2 (hACE2), allowing virus dock on cell membrane follow by viral entry. Here, we use high‐speed atomic force microscopy (HS‐AFM) for real‐time visualization of S, and its interaction with hACE2 small extracellular vesicles (sEVs). Results show conformational heterogeneity flexibility S stalk receptor‐binding domain (RBD), pH/temperature‐induced change S. in an S‐ACE2 complex appears as all‐RBD up conformation....
Abstract Protamines (PRMs) play a crucial role in sperm chromatin condensation, replacing histones to form nucleo–PRM structures, specifically PRM–DNA complexes. Despite their importance reproduction, the detailed mechanisms underlying PRM-mediated DNA condensation have remained elusive. In this study, we employed high-speed atomic force microscopy (HS-AFM) directly visualize real-time binding dynamics of PRM under physiological conditions. Our HS-AFM observations reveal that insertion...
Small extracellular vesicles (sEVs), which carry lipids, proteins and RNAs from their parent cells, serve as biomarkers for specific cell types biological states. These vesicles, including exosomes microvesicles, facilitate intercellular communication by transferring cellular components between cells. Current methods, such ultracentrifugation Tim-4 affinity method, yield high-purity sEVs. However, despite small size, purified sEVs remain heterogeneous due to varied intracellular origins. In...
Master transcription factors such as TP63 establish super-enhancers (SEs) to drive core transcriptional networks in cancer cells, yet the spatiotemporal regulation of SEs within nucleus remains unknown. The nuclear pore complex (NPC) may tether where RNA export rates are maximal. Here, we report that NUP153, a component NPC, anchors NPC and enhances expression by maximizing mRNA export. This anchoring is mediated through protein-protein interaction between intrinsically disordered regions...
DNA–histone interaction is always perturbed by epigenetic regulators to regulate gene expression. Direct visualization of this yet be achieved. By using high-speed atomic force microscopy (HS-AFM), we have observed the dynamic H2A interaction. HS-AFM movies demonstrate globular core and disordered tail H2A. DNA–H2A formed classic "beads-on-string" conformation on poly-l-lysine (PLL) lipid substrates. Notably, a short-linearized double-stranded DNA (dsDNA), resembling an inchworm, wrapped...
Small extracellular vesicles (sEVs) play a crucial role in local and distant cell communication. The intrinsic properties of sEVs make them compatible biomaterials for drug delivery, vaccines, theranostic nanoparticles. Although sEV proteomics have been robustly studied, direct instantaneous assessment structure dynamics remains difficult. Here, we use the high-speed atomic force microscopy (HS-AFM) to evaluate nanotopological changes with respect different physicochemical stresses including...
Anti-spike neutralizing antibodies (S NAbs) have been developed for prevention and treatment against COVID-19. The nanoscopic characterization of the dynamic interaction between spike proteins S NAbs remains difficult. By using high-speed atomic force microscopy (HS-AFM), we elucidate molecular property an NAb its with proteins. appeared as monomers a Y conformation at low density formed hexameric oligomers high density. NAb–spike protein RBD induces neither opening nor S1 subunit shedding....
Nuclear pore complexes (NPCs) on the nuclear membrane surface have a crucial function in controlling movement of small molecules and macromolecules between cell nucleus cytoplasm through their intricate core channel resembling spiderweb with several layers. Currently, there are few methods available to accurately measure dynamics pores membranes at nanoscale. The limitation traditional optical imaging is due diffraction, which prevents achieving required resolution for observing diverse...
Open reading frame 6 (ORF6), the accessory protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that suppresses host type-I interferon signaling, possesses amyloidogenic sequences. ORF6 peptides self-assemble to produce cytotoxic amyloid fibrils. Currently, molecular properties remain elusive. Here, we investigate structural dynamics full-length in a near-physiological environment using high-speed atomic force microscopy. oligomers were ellipsoidal and readily assembled...