Jan. 2024, 令和5年度 画像科学奨励賞, コニカミノルタ科学技術振興財団

Aug. 2022, 文科省「世界で活躍できる研究者育成事業」TRiSTARフェロー（筑波大学拠点）

Oct. 2021, 第30回ポリマー材料フォーラム 広報委員会パブリシティ賞 ポスター
稲益 礼奈、山口 央基、新井 達也、倉持 昌弘、三尾 和弘、佐々木 裕次
Others

Aug. 2019, Next Leader Award, Future of the Nematode Studies 2019

Dec. 2015, Best Presentation Award (runner-up), DAILAB CAFE-PLUS series 03
Masahiro Kuramochi

Nov. 2014, Best Presentation Award (Graduate Student Session), The Irago Conference 2014
Masahiro Kuramochi

Jun. 2011, 大学院第一種奨学金 特に優れた業績による返還免除, 日本学生支援機構
倉持 昌弘

Mar. 2011, winning of prize essay, OIST
Masahiro Kuramochi

Jan. 2011, ABA Travel Award, 7th Asia Biophysics Association (ABA) Symposium
Masahiro Kuramochi

Micro-second time-resolved X-ray single-molecule internal motions of SARS-CoV-2 spike variants
Daisuke Sasaki; Tatsuya Arai; Yue Yang; Masahiro Kuramochi; Wakako Furuyama; Asuka Nanbo; Hiroshi Sekiguchi; Nobuhiro Morone; Kazuhiro Mio; Yuji C. Sasaki, Elsevier BV
Biochemistry and Biophysics Reports, Jul. 2024, [Reviewed]

Adenosine triphosphate induces amorphous aggregation of amyloid β by increasing Aβ dynamics
Masahiro Kuramochi; Momoka Nakamura; Hiroto Takahashi; Tomoe Komoriya; Teisuke Takita; Ngan Thi Kim Pham; Kiyoshi Yasukawa; Kazuaki Yoshimune, Lead, Abstract

Amyloid β (Aβ) aggregates into two distinct fibril and amorphous forms in the brains of patients with Alzheimer’s disease. Adenosine triphosphate (ATP) is a biological hydrotrope that causes Aβ to form amorphous aggregates and inhibit fibril formation at physiological concentrations. Based on diffracted X-ray blinking (DXB) analysis, the dynamics of Aβ significantly increased immediately after ATP was added compared to those in the absence and presence of ADP and AMP, and the effect diminished after 30 min as the aggregates formed. In the presence of ATP, the β-sheet content of Aβ gradually increased from the beginning, and in the absence of ATP, the content increased rapidly after 180 min incubation, as revealed by a time-dependent thioflavin T fluorescence assay. Images of an atomic force microscope revealed that ATP induces the formation of amorphous aggregates with an average diameter of less than 100 nm, preventing fibrillar formation during 4 days of incubation at 37 °C. ATP may induce amorphous aggregation by increasing the dynamics of Aβ, and as a result, the other aggregation pathway is omitted. Our results also suggest that DXB analysis is a useful method to evaluate the inhibitory effect of fibrillar formation., Springer Science and Business Media LLC
Scientific Reports, Apr. 2024, [Reviewed]

タイヤゴム微粒子と高分子のX線観測
佐々木 裕次; 倉持 昌弘, Last
Colloid & Interface Communications, Mar. 2024, [Invited]

Real-time tilting and twisting motions of ligand-bound states of α7 nicotinic acetylcholine receptor
Yue Yang; Tatsuya Arai; Daisuke Sasaki; Masahiro Kuramochi; Hidetoshi Inagaki; Sumiko Ohashi; Hiroshi Sekiguchi; Kazuhiro Mio; Tai Kubo; Yuji C. Sasaki, Abstract

The α7 nicotinic acetylcholine receptor is a member of the nicotinic acetylcholine receptor family and is composed of five α7 subunits arranged symmetrically around a central pore. It is localized in the central nervous system and immune cells and could be a target for treating Alzheimer’s disease and schizophrenia. Acetylcholine is a ligand that opens the channel, although prolonged application rapidly decreases the response. Ivermectin was reported as one of the positive allosteric modulators, since the binding of Ivermectin to the channel enhances acetylcholine-evoked α7 currents. One research has suggested that tilting motions of the nicotinic acetylcholine receptor are responsible for channel opening and activation. To verify this hypothesis applies to α7 nicotinic acetylcholine receptor, we utilized a diffracted X-ray tracking method to monitor the stable twisting and tilting motion of nAChR α7 without a ligand, with acetylcholine, with Ivermectin, and with both of them. The results show that the α7 nicotinic acetylcholine receptor twists counterclockwise with the channel transiently opening, transitioning to a desensitized state in the presence of acetylcholine and clockwise without the channel opening in the presence of Ivermectin. We propose that the conformational transition of ACh-bound nAChR α7 may be due to the collective twisting of the five α7 subunits, resulting in the compression and movement, either downward or upward, of one or more subunits, thus manifesting tilting motions. These tilting motions possibly represent the transition from the resting state to channel opening and potentially to the desensitized state., Springer Science and Business Media LLC
European Biophysics Journal, Jan. 2024, [Reviewed]

Ubiquitination of Major Histocompatibility Complex II Changes Its Immunological Recognition Structure
Yuko Kozono; Masahiro Kuramochi; Yuji C. Sasaki; Haruo Kozono, Ubiquitination is a process that dictates the lifespan of major histocompatibility complex class II (MHC II)/peptide complexes on antigen-presenting cells. This process is tightly controlled by the levels of ubiquitin ligases, and disruptions in the turnover of MHC II can lead to the improper development of CD4+ T cells within the thymus and hinder the formation of regulatory T cells in the peripheral tissue. To investigate the underlying mechanisms, we utilized dendritic cells lacking the Membrane-associated RING-CH (MARCH) I ubiquitin ligase. We discovered that the overexpression of MARCH I decreases the interaction with LAG-3. Moreover, the MHC II molecules tethered with ubiquitin also showed diminished binding to LAG-3. We employed Diffracted X-ray Blinking (DXB), a technique used for single-molecule X-ray imaging, to observe the protein movements on live cells in real time. Our observations indicated that the normal MHC II molecules moved more rapidly across the cell surface compared to those on the MARCH I-deficient dendritic cells or MHC II KR mutants, which is likely a result of ubiquitination. These findings suggest that the signaling from ubiquitinated MHC II to the T cell receptor differs from the non-ubiquitinated forms. It appears that ubiquitinated MHC II might not be quickly internalized, but rather presents antigens to the T cells, leading to a range of significant immunological responses., MDPI AG
International Journal of Molecular Sciences, Dec. 2023, [Reviewed]

Time-Resolved X-ray Observation of Intracellular Crystallized Protein in Living Animal.
Masahiro Kuramochi; Ibuki Sugawara; Yoichi Shinkai; Kazuhiro Mio; Yuji C Sasaki, Lead, Understanding the cellular environment as molecular crowding that supports the structure-specific functional expression of biomolecules has recently attracted much attention. Time-resolved X-ray observations have the remarkable capability to capture the structural dynamics of biomolecules with subnanometre precision. Nevertheless, the measurement of the intracellular dynamics within live organisms remains a challenge. Here, we explore the potential of utilizing crystallized proteins that spontaneously form intracellular crystals to investigate their intracellular dynamics via time-resolved X-ray observations. We generated transgenic Caenorhabditis elegans specifically expressing the crystallized protein in cells and observed the formation of the protein aggregates within the animal cells. From the toxic-effect observations, the aggregates had minimal toxic effects on living animals. Fluorescence observations showed a significant suppression of the translational diffusion movements in molecules constituting the aggregates. Moreover, X-ray diffraction measurements provided diffraction signals originating from these molecules. We also observed the blinking behaviour of the diffraction spots, indicating the rotational motion of these crystals within the animal cells. A diffracted X-ray blinking (DXB) analysis estimated the rotational motion of the protein crystals on the subnanometre scale. Our results provide a time-resolved X-ray diffraction technique for the monitoring of intracellular dynamics.
International journal of molecular sciences, Nov. 2023, [Reviewed]

The ice-binding site of antifreeze protein irreversibly binds to cell surface for its hypothermic protective function
Yue Yang; Akari Yamauchi; Sakae Tsuda; Masahiro Kuramochi; Kazuhiro Mio; Yuji C. Sasaki; Tatsuya Arai
Biochemical and Biophysical Research Communications, Nov. 2023, [Reviewed]

Comparison of the Molecular Motility of Tubulin Dimeric Isoforms: Molecular Dynamics Simulations and Diffracted X-ray Tracking Study
Tsutomu Yamane; Takahiro Nakayama; Toru Ekimoto; Masao Inoue; Keigo Ikezaki; Hiroshi Sekiguchi; Masahiro Kuramochi; Yasuo Terao; Ken Judai; Minoru Saito; Mitsunori Ikeguchi; Yuji C. Sasaki, Tubulin has been recently reported to form a large family consisting of various gene isoforms; however, the differences in the molecular features of tubulin dimers composed of a combination of these isoforms remain unknown. Therefore, we attempted to elucidate the physical differences in the molecular motility of these tubulin dimers using the method of measurable pico-meter-scale molecular motility, diffracted X-ray tracking (DXT) analysis, regarding characteristic tubulin dimers, including neuronal TUBB3 and ubiquitous TUBB5. We first conducted a DXT analysis of neuronal (TUBB3-TUBA1A) and ubiquitous (TUBB5-TUBA1B) tubulin dimers and found that the molecular motility around the vertical axis of the neuronal tubulin dimer was lower than that of the ubiquitous tubulin dimer. The results of molecular dynamics (MD) simulation suggest that the difference in motility between the neuronal and ubiquitous tubulin dimers was probably caused by a change in the major contact of Gln245 in the T7 loop of TUBB from Glu11 in TUBA to Val353 in TUBB. The present study is the first report of a novel phenomenon in which the pico-meter-scale molecular motility between neuronal and ubiquitous tubulin dimers is different., MDPI AG
International Journal of Molecular Sciences, Oct. 2023, [Reviewed]

Direct observation of 890 ns dynamics of carbon black and polybutadiene in rubber materials using diffracted x-ray blinking
Masahiro Kuramochi; Henry J. Kirkwood; Jayanath C. P. Koliyadu; Romain Letrun; Raphael de Wijn; Chan Kim; Tomomi Masui; Kazuhiro Mio; Tatsuya Arai; Hiroshi Sekiguchi; Hiroyuki Kishimoto; Adrian P. Mancuso; Tokushi Sato; Yuji C. Sasaki, Lead, Dynamic behavior in soft matter physics, biology, and nanoscience frequently occurs on submicrosecond timescales. Diffracted x-ray blinking (DXB) is a unique method that can provide a broad range of spatial scale information and is becoming an attractive tool for use at high repetition rate x-ray facilities. In this study, we performed DXB experiments with 890 ns time resolution at the European X-ray Free-Electron Laser Facility to obtain dynamic information about rubber samples that are typically used in automobile tires. Time-resolved scattering was simultaneously recorded for two samples that mainly consisted of carbon black (CB) and polybutadiene (PB). These samples contained either graphitized or non-graphitized CB and displayed significantly different dynamics. A clear interaction between CB and PB was observed, indicating that the mobility of PB was changed by the introduction of CB. Restricted polymer motion was observed in the q-range of 0.78–1.58 Å−1 regions. Our results suggest that the particle network can be flexibly controlled without impairing the mechanical strength of the rubber., AIP Publishing
Applied Physics Letters, Sep. 2023, [Reviewed]

Real-Time Observation of Capsaicin-Induced Intracellular Domain Dynamics of TRPV1 Using the Diffracted X-ray Tracking Method
Kazuhiro Mio; Tatsunari Ohkubo; Daisuke Sasaki; Tatsuya Arai; Mayui Sugiura; Shoko Fujimura; Shunsuke Nozawa; Hiroshi SEKIGUCHI; Masahiro Kuramochi; Yuji C. SASAKI, The transient receptor potential vanilloid type 1 (TRPV1) is a multimodal receptor which responds to various stimuli, including capsaicin, protons, and heat. Recent advances in cryo-electron microscopy have revealed the structures of TRPV1. However, due to the large size of TRPV1 and its structural complexity, the detailed process of channel gating has not been well documented. In this study, we applied the diffracted X-ray tracking (DXT) technique to analyze the intracellular domain dynamics of the TRPV1 protein. DXT enables the capture of intramolecular motion through the analysis of trajectories of Laue spots generated from attached gold nanocrystals. Diffraction data were recorded at two different frame rates: 100 μs/frame and 12.5 ms/frame. The data from the 100 μs/frame recording were further divided into two groups based on the moving speed, using the lifetime filtering technique, and they were analyzed separately. Capsaicin increased the slope angle of the MSD curve of the C-terminus in 100 μs/frame recording, which accompanied a shifting of the rotational bias toward the counterclockwise direction, as viewed from the cytoplasmic side. This capsaicin-induced fluctuation was not observed in the 12.5 ms/frame recording, indicating that it is a high-frequency fluctuation. An intrinsiccounterclockwise twisting motion was observed in various speed components at the N-terminus, regardless of the capsaicin administration. Additionally, the competitive inhibitor AMG9810 induced a clockwise twisting motion, which is the opposite direction to capsaicin. These findings contribute to our understanding of the activation mechanisms of the TRPV1 channel.
Membranes, Jul. 2023, [Reviewed]

Unraveling the Structural and Property Differences between Highly Similar Chiral and Racemic Crystals Composed of Analogous Molecules
Kazuki Ishizaki; Daisuke Takagi; Toru Asahi; Masahiro Kuramochi; Takuya Taniguchi, American Chemical Society (ACS)
Crystal Growth & Design, Jun. 2023, [Reviewed]

The effect of ice-binding proteins on the cryopreservation of Caenorhabditis elegans
Masahiro Kuramochi; Tatsuya Arai; Kazuhiro Mio; Sakae Tsuda; Yuji C Sasaki, Lead
microPublication Biology, Apr. 2023, [Reviewed]

Observation of molecular motions in polymer thin films by laboratory grazing incidence diffracted X-ray blinking
Rena Inamasu; Hiroki Yamaguchi; Tatsuya Arai; Jaewon Chang; Masahiro Kuramochi; Kazuhiro Mio; Yuji C. Sasaki, Abstract

Research on polymer surfaces has shown that the mobilities of polymer chains, which affect the aggregation state and thus the physical properties of the material, differ between the surface and bulk. However, the mobilities of the surface polymers have not been fully characterized. Therefore, we propose a time-resolved method for evaluating surface mobility. This measurement scheme is called grazing incidence diffracted X-ray blinking (GI-DXB) and can be used to evaluate the molecular motions occurring at polymer surfaces by continuously measuring X-ray diffraction patterns near the total reflection angle over small time periods. In this study, the crystallized polymer poly{2-(perfluorooctyl)ethyl acrylate}(PC₈FA) was measured. The decay constants, which are indexes of molecular motions, were calculated to be 3.98 × 10⁻³s⁻¹ for the fluoroalkyl groups in the side chains observed along the in-plane direction and 3.36 × 10⁻³s⁻¹ for the lamellar structure observed along the out-of-plane direction when 2000 diffraction profiles of 500 ms were recorded and the incident angle was 0.07°. In contrast, transmission DXB indicated decay constants of 2.63 × 10⁻³s⁻¹ for the side chains and 2.87 × 10⁻³s⁻¹ for the lamellar structures. These results suggested that the PC₈FA surface is mobile, because a larger decay constant indicates a higher mobility. GI-DXB can be used to measure surface dynamics. The authors contend that GI-DXB is a highly versatile tool because it allows the evaluation of local motions with a laboratory X-ray system, and these motions cannot be detected by conventional surface analyses. This measurement scheme may facilitate the development of high-performance polymers and discovery of new physical properties., Springer Science and Business Media LLC
Polymer Journal, Feb. 2023, [Reviewed]

Visualizing Intramolecular Dynamics of Membrane Proteins
Tatsunari Ohkubo; Takaaki Shiina; Kayoko Kawaguchi; Daisuke Sasaki; Rena Inamasu; Yue Yang; Zhuoqi Li; Keizaburo Taninaka; Masaki Sakaguchi; Shoko Fujimura; Hiroshi Sekiguchi; Masahiro Kuramochi; Tatsuya Arai; Sakae Tsuda; Yuji C. Sasaki; Kazuhiro Mio, Membrane proteins play important roles in biological functions, with accompanying allosteric structure changes. Understanding intramolecular dynamics helps elucidate catalytic mechanisms and develop new drugs. In contrast to the various technologies for structural analysis, methods for analyzing intramolecular dynamics are limited. Single-molecule measurements using optical microscopy have been widely used for kinetic analysis. Recently, improvements in detectors and image analysis technology have made it possible to use single-molecule determination methods using X-rays and electron beams, such as diffracted X-ray tracking (DXT), X-ray free electron laser (XFEL) imaging, and cryo-electron microscopy (cryo-EM). High-speed atomic force microscopy (HS-AFM) is a scanning probe microscope that can capture the structural dynamics of biomolecules in real time at the single-molecule level. Time-resolved techniques also facilitate an understanding of real-time intramolecular processes during chemical reactions. In this review, recent advances in membrane protein dynamics visualization techniques were presented., MDPI AG
International Journal of Molecular Sciences, Nov. 2022, [Reviewed]

A mutation to a fish ice-binding protein synthesized in transgenic Caenorhabditis elegans modulates its cold tolerance.
Masahiro Kuramochi; Shumiao Zhu; Chiaki Takanashi; Yue Yang; Tatsuya Arai; Yoichi Shinkai; Motomichi Doi; Kazuhiro Mio; Sakae Tsuda; Yuji C Sasaki, A cryoprotectant known as ice-binding protein (IBP) is thought to facilitate the cold survival of plants, insects, and fungi. Here, we prepared a genetically modified Caenorhabditis elegans strain to synthesize fish-derived IBPs in its body wall muscles and examined whether the antifreeze activity modification of this IBP by point mutation affects the cold tolerance of this worm. We chose a 65-residue IBP identified from notched-fin eelpout, for which the replacement of the 20th alanine residue (A20) modifies its antifreeze activity. These mutant proteins are denoted A20L, A20G, A20T, A20V, and A20I along with the wild-type (WT) protein. We evaluated the survival rate (%) of the transgenic C. elegans that synthesized each IBP mutant following 24 h of preservation at -5, +2, and +5 °C. Significantly, a dramatic improvement in the survival rate was detected for the worms synthesizing the activity-enhanced mutants (A20T and A20I), especially at +2 °C. In contrast, the rate was not improved by the expression of the defective mutants (A20L, A20G, WT and A20V). The survival rate (%) probably correlates with the antifreeze activity of the IBP. These data suggest that IBP protects the cell membrane by employing its ice-binding mechanism, which ultimately improves the cold tolerance of an IBP-containing animal.
Biochemical and biophysical research communications, Nov. 2022

Dynamic observations of various oligomers in amyloid β isoforms using laboratory diffracted X-ray blinking
Jaewon Chang; Tatsuya Arai; Masahiro Kuramochi; Rena Inamasu; Zhuoqi Lee; Tatsunari Ohkubo; Kazuhiro Mio; Yuji C. Sasaki, Acceleration of societal ageing has increased the global incidence of geriatric diseases such as Alzheimer's disease (AD), and the demands for proper diagnosis and monitoring of those diseases are also increasing daily. We utilized diffracted X-ray blinking (DXB) for amyloid β (Aβ) isoforms, which are thought to be closely related to AD, to discriminate among the dynamics of individual particles in early and long-term oligomerisation and aggregation inhibiting environments. Among the various Aβ isoforms, the dynamics of Aβ (1-42), which is known to be the most toxic form, were the slowest (the dynamics were lower by 78% com-pared with short-term incubation), and the dynamics were restored (the dynamics increased by 105% compared with normal aggregation) in an environment that suppressed oligomerisation of Aβ (1-42). It has been confirmed that the use of DXB allows measurements of dynamics related to the functional states of the target molecules., Elsevier BV
Biochemistry and Biophysics Reports, Sep. 2022, [Reviewed]

Diffracted X-ray Tracking Method for Measuring Intramolecular Dynamics of Membrane Proteins
Shoko Fujimura; Kazuhiro Mio; Tatsunari Ohkubo; Tatsuya Arai; Masahiro Kuramochi; Hiroshi Sekiguchi; Yuji C. Sasaki, Membrane proteins change their conformations in response to chemical and physical stimuli and transmit extracellular signals inside cells. Several approaches have been developed for solving the structures of proteins. However, few techniques can monitor real-time protein dynamics. The diffracted X-ray tracking method (DXT) is an X-ray-based single-molecule technique that monitors the internal motion of biomolecules in an aqueous solution. DXT analyzes trajectories of Laue spots generated from the attached gold nanocrystals with a two-dimensional axis by tilting (θ) and twisting (χ). Furthermore, high-intensity X-rays from synchrotron radiation facilities enable measurements with microsecond-timescale and picometer-spatial-scale intramolecular information. The technique has been applied to various membrane proteins due to its superior spatiotemporal resolution. In this review, we introduce basic principles of DXT, reviewing its recent and extended applications to membrane proteins and living cells, respectively., MDPI AG
International Journal of Molecular Sciences, Feb. 2022

Dynamic motions of ice-binding proteins in living Caenorhabditis elegans using diffracted X-ray blinking and tracking
Masahiro Kuramochi; Yige Dong; Yue Yang; Tatsuya Arai; Rio Okada; Yoichi Shinkai; Motomichi Doi; Kouki Aoyama; Hiroshi Sekiguchi; Kazuhiro Mio; Sakae Tsuda; Yuji C. Sasaki, Lead, The dynamic properties of protein molecules are involved in the relationship between their structure and function. Time-resolved X-ray observation enables capturing the structures of biomolecules with picometre-scale precision. However, this technique has yet to be implemented in living animals. Here, we examined diffracted X-ray blinking (DXB) and diffracted X-ray tracking (DXT) to observe the dynamics of a protein located on intestinal cells in adult Caenorhabditis elegans. This in vivo tissue-specific DXB was examined at temperatures from 20 °C to -10 °C for a recombinant ice-binding protein from Antarctomyces psychrotrophicus (AnpIBP) connected with the cells through a transmembrane CD4 protein equipped with a glycine-serine linker. AnpIBP inhibits ice growth at subzero temperatures by binding to ice crystals. We found that the rotational motion of AnpIBP decreases at -10 °C. In contrast, the motion of the AnpIBP mutant, which has a defective ice-binding ability, did not decrease at -10 °C. The twisting and tilting motional speeds of AnpIBPs measured above 5 °C by DXT were always higher than those of the defective AnpIBP mutant. These results suggest that wild-type AnpIBP is highly mobile in solution, and it is halted at subzero temperatures through ice binding. DXB and DXT allow for exploring protein behaviour in live animals with subnano resolution precision., Elsevier BV
Biochemistry and Biophysics Reports, Feb. 2022, [Reviewed]

Superelasticity of a photo-actuating chiral salicylideneamine crystal
Takuya Taniguchi; Kazuki Ishizaki; Daisuke Takagi; Kazuki Nishimura; Hiroki Shigemune; Masahiro Kuramochi; Yuji C. Sasaki; Hideko Koshima; Toru Asahi, Superelasticity is a type of elastic response to an applied external force, caused by a phase transformation. Actuation of materials is also an elastic response to external stimuli such as light and heat. Although both superelasticity and actuation are deformations resulting from stimulus-induced stress, there is a phenomenological difference between the two with respect to whether force is an input or an output. Here, we report that a molecular crystal manifests superelasticity during photo-actuation under light irradiation. The crystal exhibits stepwise twisted actuation due to two effects, photoisomerization and photo-triggered phase transition, and the actuation behavior is simulated based on a dynamic multi-layer model. The simulation, in turn, reveals how the photoisomerization and phase transition progress in the crystal, indicating superelasticity induced by modest stress due to the formation of photoproducts. This work provides not only a successful simulation of stepwise twisted actuation, but also to the best of our knowledge the first indication of superelasticity induced by light., Springer Science and Business Media LLC
Communications Chemistry, Jan. 2022, [Reviewed]

X線1分子追跡法を用いたTRPイオンチャネル運動の高速時分割計測と制御機構の理解
三尾 和弘; 藤村 章子; 倉持 昌弘; 関口 博史; 佐々木 裕次, 日本放射光学会
放射光 : 日本放射光学会誌 = Journal of the Japanese Society for Synchrotron Radiation Research, Sep. 2021

New Family Members of FG Repeat Proteins and Their Unexplored Roles During Phase Separation
Yoichi Shinkai; Masahiro Kuramochi; Takamitsu Miyafusa, Corresponding, The condensation and compartmentalization of biomacromolecules in the cell are driven by the process of phase separation. The main effectors of phase separation are intrinsically disordered proteins, which include proteins with a phenylalanine-glycine (FG) repeat domain. Our understanding of the biological function of FG repeat proteins during phase separation has been mainly derived from recent research on a member of the nuclear pore complex proteins, nucleoporins containing FG repeat domain (FG-NUPs). FG-NUPs form meshwork structures by inter- and intra-molecular FG domain interactions, which confine the nucleo-cytoplasmic exchange. Whereas FG-NUPs localize in the nuclear membrane, other FG repeat proteins reside in the cytoplasm and the nucleoplasm, and the biological function of the FG repeat domain of these proteins is not well described. In the present review, we list the FG repeat proteins that are known to phase separate in the cell, and review their biological functions. We extract the unraveled features of FG repeat proteins as an activator of barrier formation and homotypic cell-cell interactions. Understanding the regulatory mechanisms of FG repeat proteins will provide a potential delivery tool for therapeutic reagents., Frontiers Media SA
Frontiers in Cell and Developmental Biology, Jul. 2021, [Reviewed]

Laboratory diffracted x-ray blinking to monitor picometer motions of protein molecules and application to crystalline materials
Tatsuya Arai; Rena Inamasu; Hiroki Yamaguchi; Daisuke Sasaki; Ayana Sato-Tomita; Hiroshi Sekiguchi; Kazuhiro Mio; Sakae Tsuda; Masahiro Kuramochi; Yuji C. Sasaki, Corresponding, In recent years, real-time observations of molecules have been required to understand their behavior and function. To date, we have reported two different time-resolved observation methods: diffracted x-ray tracking and diffracted x-ray blinking (DXB). The former monitors the motion of diffracted spots derived from nanocrystals labeled onto target molecules, and the latter measures the fluctuation of the diffraction intensity that is highly correlated with the target molecular motion. However, these reports use a synchrotron x-ray source because of its high average flux, resulting in a high time resolution. Here, we used a laboratory x-ray source and DXB to measure the internal molecular dynamics of three different systems. The samples studied were bovine serum albumin (BSA) pinned onto a substrate, antifreeze protein (AFP) crystallized as a single crystal, and poly{2-(perfluorooctyl)ethyl acrylate} (PC8FA) polymer between polyimide sheets. It was found that not only BSA but also AFP and PC8FA molecules move in the systems. In addition, the molecular motion of AFP molecules was observed to increase with decreasing temperature. The rotational diffusion coefficients (DR) of BSA, AFP, and PC8FA were estimated to be 0.73 pm2/s, 0.65 pm2/s, and 3.29 pm2/s, respectively. Surprisingly, the DR of the PC8FA polymer was found to be the highest among the three samples. This is the first report that measures the molecular motion of a single protein crystal and polymer by using DXB with a laboratory x-ray source. This technique can be applied to any kind of crystal and crystalline polymer and provides atomic-order molecular information., AIP Publishing
Structural Dynamics, Jul. 2021, [Reviewed]

Structural basis for the coiled-coil architecture of human CtIP
C. R. Morton; N. J. Rzechorzek; J. D. Maman; M. Kuramochi; H. Sekiguchi; R. Rambo; Y. C. Sasaki; O. R. Davies; L. Pellegrini, The DNA repair factor CtIP has a critical function in double-strand break (DSB) repair by homologous recombination, promoting the assembly of the repair apparatus at DNA ends and participating in DNA-end resection. However, the molecular mechanisms of CtIP function in DSB repair remain unclear. Here, we present an atomic model for the three-dimensional architecture of human CtIP, derived from a multi-disciplinary approach that includes X-ray crystallography, small-angle X-ray scattering (SAXS) and diffracted X-ray tracking (DXT). Our data show that CtIP adopts an extended dimer-of-dimers structure, in agreement with a role in bridging distant sites on chromosomal DNA during the recombinational repair. The zinc-binding motif in the CtIP N-terminus alters dynamically the coiled-coil structure, with functional implications for the long-range interactions of CtIP with DNA. Our results provide a structural basis for the three-dimensional arrangement of chains in the CtIP tetramer, a key aspect of CtIP function in DNA DSB repair., The Royal Society
Open Biology, Jun. 2021, [Reviewed]

Diffracted X-ray blinking measurements of interleukin 15 receptors in the inner/outer membrane of living NK cells
Jaewon Chang; Yonugseok Baek; Injee Lee; Hiroshi Sekiguchi; Kouhei Ichiyanagi; Kazuhiro Mio; Shunsuke Nozawa; Ryo Fukaya; Shin-ichi Adachi; Masahiro Kuramochi; Yuji C. Sasaki, Corresponding, Interleukin 15 receptor (IL-15R) is a transmembrane signalling protein consisting of 3 subsets: α, β (IL-15Rβ), and γ (γ ). IL-2 and IL-15 share the signalling domains IL-15Rβ and γ , although they bind to intrinsic α-subsets and non-signalling domains. Additionally, IL-2 and IL-15 play different roles; therefore, there have been many observations of the dynamic behaviours of IL-15R, which are linked to physiological functions. For more practical discrimination between IL-2 and IL-15, a study was designed and carried out in which α-subsets were removed and a cytoplasmic inhibitor was applied to create a simplified environment in which secondary signalling molecules were reduced. We also applied a new measurement method, diffracted X-ray blinking (DXB), to achieve higher accuracy (<0.01 Å). The dynamics of IL-2 binding (confined motion, max range = 0.71 Å) and IL-15 binding (normal motion) in live natural killer cells were different. We also confirmed. that DXB was a suitable method to quantitatively evaluate the transmembrane protein dynamics of inner/outer live cell membranes by labeling the extracellular domain since the measurements were dependent on the cytosolic environment. c c, Elsevier BV
Biochemical and Biophysical Research Communications, Jun. 2021, [Reviewed]

Living-Cell Diffracted X-ray Tracking Analysis Confirmed Internal Salt Bridge Is Critical for Ligand-Induced Twisting Motion of Serotonin Receptors
Kazuhiro Mio; Shoko Fujimura; Masaki Ishihara; Masahiro Kuramochi; Hiroshi Sekiguchi; Tai Kubo; Yuji C. Sasaki, Serotonin receptors play important roles in neuronal excitation, emotion, platelet aggregation, and vasoconstriction. The serotonin receptor subtype 2A (5-HT2AR) is a Gq-coupled GPCR, which activate phospholipase C. Although the structures and functions of 5-HT2ARs have been well studied, little has been known about their real-time dynamics. In this study, we analyzed the intramolecular motion of the 5-HT2AR in living cells using the diffracted X-ray tracking (DXT) technique. The DXT is a very precise single-molecular analytical technique, which tracks diffraction spots from the gold nanocrystals labeled on the protein surface. Trajectory analysis provides insight into protein dynamics. The 5-HT2ARs were transiently expressed in HEK 293 cells, and the gold nanocrystals were attached to the N-terminal introduced FLAG-tag via anti-FLAG antibodies. The motions were recorded with a frame rate of 100 μs per frame. A lifetime filtering technique demonstrated that the unliganded receptors contain high mobility population with clockwise twisting. This rotation was, however, abolished by either a full agonist α-methylserotonin or an inverse agonist ketanserin. Mutation analysis revealed that the “ionic lock” between the DRY motif in the third transmembrane segment and a negatively charged residue of the sixth transmembrane segment is essential for the torsional motion at the N-terminus of the receptor., MDPI AG
International Journal of Molecular Sciences, May 2021, [Reviewed]

Tilting and rotational motions of silver halide crystal with diffracted X-ray blinking
Masahiro Kuramochi; Hiroki Omata; Masaki Ishihara; Sander Ø. Hanslin; Masaichiro Mizumaki; Naomi Kawamura; Hitoshi Osawa; Motohiro Suzuki; Kazuhiro Mio; Hiroshi Sekiguchi; Yuji C. Sasaki, Lead, The dynamic properties of crystalline materials are important for understanding their local environment or individual single-grain motions. A new time-resolved observation method is required for use in many fields of investigation. Here, we developed in situ diffracted X-ray blinking to monitor high-resolution diffraction patterns from single-crystal grains with a 50 ms time resolution. The diffraction spots of single grains of silver halides and silver moved in the θ and χ directions during the photolysis chemical reaction. The movements of the spots represent tilting and rotational motions. The time trajectory of the diffraction intensity reflecting those motions was analysed by using single-pixel autocorrelation function (sp-ACF). Single-pixel ACF analysis revealed significant differences in the distributions of the ACF decay constants between silver halides, suggesting that the motions of single grains are different between them. The rotational diffusion coefficients for silver halides were estimated to be accurate at the level of approximately 0.1 to 0.3 pm²/s. Furthermore, newly formed silver grains on silver halides correlated with their ACF decay constants. Our high-resolution atomic scale measurement—sp-ACF analysis of diffraction patterns of individual grains—is useful for evaluating physical properties that are broadly applicable in physics, chemistry, and materials science., Springer Science and Business Media LLC
Scientific Reports, Mar. 2021, [Reviewed]

Agonist and Antagonist-Diverted Twisting Motions of a Single TRPV1 Channel
Shoko Fujimura; Kazuhiro Mio; Masahiro Kuramochi; Hiroshi Sekiguchi; Keigo Ikezaki; Muneyo Mio; Kowit Hengphasatporn; Yasuteru Shigeta; Tai Kubo; Yuji C. Sasaki, Transient receptor potential vanilloid type 1 (TRPV1) channels are activated by heat, vanilloids, and extracellular protons. Cryo-EM has revealed various conformations of TRPV1, and these structures suggest an intramolecular twisting motion in response to ligand binding. However, limited experimental data support this observation. Here, we analyzed the intramolecular motion of TRPV1 using diffracted X-ray tracking (DXT). DXT analyzes trajectories of Laue spots generated from attached gold nanocrystals and provides picometer spatial and microsecond time scale information about the intramolecular motion. We observed that both an agonist and a competitive antagonist evoked a rotating bias in TRPV1, though these biases were in opposing directions. Furthermore, the rotational bias generated by capsaicin was reversed between the wild-type and the capsaicin-insensitive Y511A mutant. Our findings bolster the understanding of the mechanisms used for activation and modulation of TRP channels, and this knowledge can be exploited for pharmacological usage such as inhibitor design., American Chemical Society (ACS)
The Journal of Physical Chemistry B, Dec. 2020, [Reviewed]

X-ray-based living-cell motion analysis of individual serotonin receptors
Kazuhiro Mio; Masaki Ishihara; Shoko Fujimura; Daisuke Sasaki; Shunsuke Nozawa; Kohei Ichiyanagi; Ryo Fukaya; Shin-ichi Adachi; Masahiro Kuramochi; Hiroshi Sekiguchi; Tai Kubo; Yuji C. Sasaki, G protein-coupled receptors (GPCRs) are seven-transmembrane proteins, which transmit extracellular signals inside cells via activating G proteins. GPCRs are involved in a wide variety of physiological functions, such as signal sensing, immune system processes, and neurotransmission. Although the structures and functions of GPCRs have been well studied, little has been known about their real-time dynamics on live cells. In this study, we used Diffracted X-ray Tracking (DXT) and Diffracted X-ray Blinking (DXB) techniques for analysis. These methods are very precise single-molecular analytical techniques that elucidate protein dynamics by analyzing the diffraction spots from the gold nanocrystals labeled on the protein surface. DXT tracks diffraction spot movements, whereas DXB analyzes continuation of signals by calculating the autocorrelation function of each pixel from the recorded data. Serotonin receptor subtype 2A (5-HT2A receptors) were transiently expressed on HEK 293 cells, and the gold nanocrystals were attached to the N-terminally introduced FLAG-tag via anti-FLAG antibodies. Fast- and mid-range motions were recorded by DXT with 100μs and 1.25 ms/frame rate, respectively. Slow-range motion was obtained using the DXB method with 100 ms/frame rate. An agonist interestingly suppressed the fluctuations of 5-HT2A receptors at the microsecond-ranged fast measurement. On the contrary, the motion was enhanced by the agonist in the hundred-millisecond-ranged slow time scale. These dual-natured data may suggest that we succeeded in extracting different modes of receptor's motion on live cells; microsecond ranged fluctuation on the cell membrane, and millisecond-ranged dynamic movement comprising interactions with intracellular signaling molecules., Elsevier BV
Biochemical and Biophysical Research Communications, Aug. 2020, [Reviewed]

Expression of Ice-Binding Proteins in Caenorhabditis elegans Improves the Survival Rate upon Cold Shock and during Freezing
Masahiro Kuramochi; Chiaki Takanashi; Akari Yamauchi; Motomichi Doi; Kazuhiro Mio; Sakae Tsuda; Yuji C. Sasaki, Lead, Ice-binding proteins (IBPs) are capable of binding ice crystals and inhibiting their growth at freezing temperatures. IBPs are also thought to stabilize the cell membrane at non-freezing temperatures near 0 °C. These two effects have been assumed to reduce cold- and freezing-induced damage to cells and tissues. However, knowledge regarding the effects of IBP on the living animals is limited. Here, we characterized the relationship between the IBP effects and the physiological role by using the nematode Caenorhabditis elegans. The expression of fish (NfeIBPs)- and fungus-derived IBPs (AnpIBPs and TisIBP8) in C. elegans improved its survival rate during exposure to 0 and -2 °C (cold shock) and -5 °C (freezing). The observed cold tolerance of C. elegans after cold shock is attributable to the stabilization of cell-membrane lipids with IBPs, and the freezing tolerance at -5 °C can be attributed to the inhibition of ice-crystal growth by the IBPs. Significantly, the survival rate of C. elegans at -5 °C was improved by expression of wild-type AnpIBP and maximized by that of TisIBP8, whereas it was lowered when a defective AnpIBP mutant was expressed. These results suggest that the ice-binding ability of IBP has a good correlation with the survival rate of C. elegans during freezing., Springer Science and Business Media LLC
Scientific Reports, Dec. 2019, [Reviewed]

An Excitatory/Inhibitory Switch From Asymmetric Sensory Neurons Defines Postsynaptic Tuning for a Rapid Response to NaCl in Caenorhabditis elegans
Masahiro Kuramochi; Motomichi Doi, Lead, Frontiers Media SA
Frontiers in Molecular Neuroscience, Jan. 2019, [Reviewed]

Toward understanding of internal motion measurement with quantum probe and cryo-EM
K. Mio; S. Fujimura; M. Kuramochi; M. Ishihara; M. Honda; M. Mio; T. Kubo; H. Sekiguchi; Y. C. Sasaki, Pesticide Science Society of Japan
Japanese Journal of Pesticide Science, 2019, [Reviewed]

線虫Ｃ．エレンガンスを用いた不凍タンパク質のin vivo解析　　　　　　　　　　　　　　　
倉持昌弘; 高梨千晶; 山内彩加林; 戸井基道; 三尾和弘; 津田栄; 佐々木裕次, Lead
低温生物工学会誌, 2019, [Reviewed]

Diffracted X-ray Blinking Tracks Single Protein Motions
Hiroshi Sekiguchi; Masahiro Kuramochi; Keigo Ikezaki; Yu Okamura; Kazuki Yoshimura; Ken Matsubara; Jae-Won Chang; Noboru Ohta; Tai Kubo; Kazuhiro Mio; Yoshio Suzuki; Leonard M. G. Chavas; Yuji C. Sasaki, Single molecule dynamics studies have begun to use quantum probes. Single particle analysis using cryo-transmission electron microscopy has dramatically improved the resolution when studying protein structures and is shifting towards molecular motion observations. X-ray free-electron lasers are also being explored as routes for determining single molecule structures of biological entities. Here, we propose a new X-ray single molecule technology that allows observation of molecular internal motion over long time scales, ranging from milliseconds up to 10(3) seconds. Our method uses both low-dose monochromatic X-rays and nanocrystal labelling technology. During monochromatic X-ray diffraction experiments, the intensity of X-ray diffraction from moving single nanocrystals appears to blink because of Brownian motion in aqueous solutions. X-ray diffraction spots from moving nanocrystals were observed to cycle in and out of the Bragg condition. Consequently, the internal motions of a protein molecule labelled with nanocrystals could be extracted from the time trajectory using this diffracted X-ray blinking (DXB) approach. Finally, we succeeded in distinguishing the degree of fluctuation motions of an individual acetylcholine-binding protein (AChBP) interacting with acetylcholine (ACh) using a laboratory X-ray source., Springer Science and Business Media LLC
Scientific Reports, Dec. 2018, [Reviewed]

Regulation of chromatin states and gene expression during HSN neuronal maturation is mediated by EOR-1/PLZF, MAU-2/cohesin loader, and SWI/SNF complex
Yoichi Shinkai; Masahiro Kuramochi; Motomichi Doi, Springer Science and Business Media LLC
Scientific Reports, Dec. 2018, [Reviewed]

X-ray observations of single bio-supramolecular photochirogenesis
Jaewon Chang; Masaki Nishijima; Hiroshi Sekiguchi; Kohei Ichiyanagi; Masahiro Kuramochi; Yoshihisa Inoue; Yuji C. Sasaki, The binding and photochirogenic behaviour of 2-anthracenecarboxylate (AC) with human serum albumin (HSA) have hitherto been investigated and comprehended as time-averaged statistical events by spectroscopic examinations and product analyses. In this study, we employed a diffracted X-ray tracking (DXT) technique to visualize the single-molecular dynamics of free and AC-loaded HSA (AC:HSA = 0, 1, 5 and 10), as well as the AC-HSA complex under photoirradiation, all of which were tethered to gold nanocrystals and hence traceable in real time by DXT. This enabled us to draw a more dynamic picture of the bio-supramolecular photochirogenesis at a single-molecule resolution, detailing the softening and flexibility enhancement of HSA upon binding of ACs to its inter-subdomain IIA-IIB site and the dynamic extrusion of AC dimers produced upon photoirradiation., Elsevier BV
Biophysical Chemistry, Nov. 2018, [Reviewed]

A Simple Method for Visualization of Locus-Specific H4K20me1 Modifications in Living Caenorhabditis elegans Single Cells
Yoichi Shinkai; Masahiro Kuramochi; Motomichi Doi,
Recently, advances in next-generation sequencing technologies have enabled genome-wide analyses of epigenetic modifications; however, it remains difficult to analyze the states of histone modifications at a single-cell resolution in living multicellular organisms because of the heterogeneity within cellular populations. Here we describe a simple method to visualize histone modifications on the specific sequence of target locus at a single-cell resolution in living Caenorhabditis elegans, by combining the LacO/LacI system and a genetically-encoded H4K20me1-specific probe, “mintbody”. We demonstrate that Venus-labeled mintbody and mTurquoise2-labeled LacI can co-localize on an artificial chromosome carrying both the target locus and LacO sequences, where H4K20me1 marks the target locus. We demonstrate that our visualization method can precisely detect H4K20me1 depositions on the her-1 gene sequences on the artificial chromosome, to which the dosage compensation complex binds to regulate sex determination. The degree of H4K20me1 deposition on the her-1 sequences on the artificial chromosome correlated strongly with sex, suggesting that, using the artificial chromosome, this method can reflect context-dependent changes of H4K20me1 on endogenous genomes. Furthermore, we demonstrate live imaging of H4K20me1 depositions on the artificial chromosome. Combined with ChIP assays, this mintbody-LacO/LacI visualization method will enable analysis of developmental and context-dependent alterations of locus-specific histone modifications in specific cells and elucidation of the underlying molecular mechanisms., Oxford University Press (OUP)
G3 Genes|Genomes|Genetics, Jul. 2018, [Reviewed]

A Computational Model Based on Multi-Regional Calcium Imaging Represents the Spatio-Temporal Dynamics in a Caenorhabditis elegans Sensory Neuron
Masahiro Kuramochi; Motomichi Doi, Lead, Public Library of Science (PLoS)
PLOS ONE, Jan. 2017, [Reviewed]

Quantitative Modeling of Neuronal Dynamics in C. elegans
Masahiro Kuramochi; Yuishi Iwasaki, Lead, Springer Berlin Heidelberg
Neural Information Processing. Theory and Algorithms, 2010, [Reviewed]

細胞内液-液相分離の攪乱とその生体応答
丸山迪代; 三村真大; 三村真大; 新木和孝; 井上明俊; 倉持昌弘; 戸井基道; 草田裕之; 冨田峻介; 新海陽一
日本生化学会大会(Web), 2023

生きた線虫の1分子動態と細胞応答を同時取得する時分割X線回折/細胞イメージング法
倉持昌弘
中谷医工計測技術振興財団年報, 2021
Lead

Dynamic Internal Motion of GPCR on Live Cells
Masaki Ishihara; Shoko Fujimura; Kohei Ichiyanagi; Shunsuke Nozawa; Shinichi Adachi; Ryo Fukaya; Masahiro Kuramochi; Hiroshi Sekiguchi; Kazuhiro Mio; Yuji C. Sasaki
BIOPHYSICAL JOURNAL, Feb. 2020, [Reviewed]

Novel In Vivo Observations of Single Protein Motions Using Laboratory X-Ray Source
Sasaki Y. C; Kuramochi M; Sekiguchi H; Mio K
Biophysical Journal, 2018, [Reviewed]

Functional Analysis of Antifreeze Proteins for Cold Tolerance Behavior and X-Ray Single Molecule Observations in C-elegans
Kuramochi Masahiro; Takanashi Chiaki; Sekiguchi Hiroshi; Doi Motomichi; Tsuda Sakae; Sasaki Yuji C
Biophysical Journal, 2018, [Reviewed]
Lead

線虫C. elegansの味覚神経活動の時空間動態解析　　　　　　　　　　　　　　　
倉持 昌弘; 戸井 基道
電子情報通信学会技術研究報告. NC, ニューロコンピューティング, Mar. 2015
Lead

Acquiring freezing and cold tolerance: The role of structure-specific functions in Ice-Binding Protein molecules　　　　　　　　　　　　　　　
倉持昌弘; 新井達也; 三尾和弘; 津田栄; 佐々木裕次
Sep. 2023

Ice-Binding Proteins in maintaining biological activity in frozen and sub-zero environments　　　　　　　　　　　　　　　
倉持昌弘; 新井達也; 三尾和弘; 津田栄; 佐々木裕次
May 2023

氷結合タンパク質は個体生物の凍結・非凍結低温環境における生存率を改善する　　　　　　　　　　　　　　　
倉持昌弘; 新井達也; 三尾和弘; 津田栄; 佐々木裕次
Dec. 2022

個体生物の凍結・低温耐性を改善する不凍タンパク質の生体内効果　　　　　　　　　　　　　　　
倉持 昌弘; 三尾 和弘; 津田 栄; 佐々木 裕次
May 2021

線虫の凍結・低温耐性を改善する氷結合タンパク質の生体内作用機序の解明　　　　　　　　　　　　　　　
倉持 昌弘; 三尾 和弘; 津田 栄; 佐々木 裕次
Apr. 2021

生きた線虫C. elegans細胞内における蛍光タンパク質結晶のX線観察 Dynamical X-ray observation of the crystallized protein in the living Caenorhabditis elegans cells　　　　　　　　　　　　　　　
菅原生蕗; 新海陽一; 倉持昌弘
2023年度量子ビームサイエンスフェスタ, Mar. 2024

サブテラヘルツ光照射による水和ダイナミクスのX線動態計測と線虫胚発生への非熱作用 Hydration Dynamics and Non-Thermal Effects on the C. elegans Embryogenesis by sub-THz Irradiation　　　　　　　　　　　　　　　
栗山稜平; 今清水正彦; 倉持昌弘
2023年度量子ビームサイエンスフェスタ, Mar. 2024

薬剤投与に対するアミロイドβのX線動態解析 Dynamic X-ray Observation of Amyloid-β Behaviour Following Drug Administration　　　　　　　　　　　　　　　
髙橋大翔; 吉宗一晃; 倉持昌弘
2023年度量子ビームサイエンスフェスタ, Mar. 2024

機械学習を用いたX線透過動画解析による未知動態探索 Searching for unknown dynamics by analysing X-ray transmission videos using machine learning　　　　　　　　　　　　　　　
倉持昌弘; 赤穂昭太郎; 佐々木裕次
2023年度量子ビームサイエンスフェスタ, Mar. 2024

凍結低温制御分子の構造特異機能の解明および個体丸ごと保存技術の開発　　　　　　　　　　　　　　　
倉持昌弘
ACT-X「環境とバイオテクノロジー」第5回領域会議, Jan. 2024

凍結低温制御分子の機能と応用　　　　　　　　　　　　　　　
倉持昌弘
TRiSTAR共創リレーセミナー, Jan. 2024

氷晶結合タンパク質分子による個体動物の凍結保存効果　　　　　　　　　　　　　　　
下谷 祐平; 倉持 昌弘
第34回日本化学会関東支部茨城地区研究交流会, Dec. 2023

氷結合タンパク質による個体動物の低温耐性獲得メカニズム　　　　　　　　　　　　　　　
中村咲; 新海陽一; 倉持昌弘
第34回日本化学会関東支部茨城地区研究交流会, Dec. 2023

氷晶結合タンパク質分子を導入した線虫Ｃ．エレガンスの乾燥耐性　　　　　　　　　　　　　　　
下瀬大輝; 倉持昌弘
第34回日本化学会関東支部茨城地区研究交流会, Dec. 2023

薬剤投与に対するアミロイドβのX線動態解析　　　　　　　　　　　　　　　
高橋大翔; 吉宗 一晃; 倉持昌弘
第34回日本化学会関東支部茨城地区研究交流会, Dec. 2023

Rotational dynamics of nanoparticles in polymer nanocomposites observed with diffracted X-ray blinking　　　　　　　　　　　　　　　
Tatsuya Arai; Masahiro Kuramochi; Kazuhiro Mio; Hiroshi Sekiguchi; Yuji C. Sasaki
MRM2023/IUMRS-ICA2023, Dec. 2023

Dynamical properties of polymer materials from X-ray transmission video using the machine learning　　　　　　　　　　　　　　　
M. Kuramochi; S. Akaho; Y.C. Sasaki
MRM2023/IUMRS-ICA2023, Dec. 2023

"氷晶結合タンパク質分子を導入した線虫Ｃ．エレガンスの乾燥耐性 Desiccation tolerance of the Caenorhabditis elegans expressing the ice-binding protein "　　　　　　　　　　　　　　　
D. Shimose; M. Kuramochi
第46回日本分子生物学会年会, Dec. 2023

Molecular mechanisms of ice-binding proteins for the cold tolerance at the unfrozen low temperature　　　　　　　　　　　　　　　
S. Nakamura; Y. Shinkai; M. Kuramochi
第46回日本分子生物学会年会, Dec. 2023

薬剤投与に対するアミロイドβのX線動態解析　　　　　　　　　　　　　　　
H. Takahashi; K. Yoshimune; M. Kuramochi
第46回日本分子生物学会年会, Dec. 2023

サブテラヘルツ光照射による水和ダイナミクスと線虫胚発生への非熱的作用 Hydration Dynamics and Non-Thermal Effects on C. elegans Embryogenesis by Sub-Terahertz Light Irradiation　　　　　　　　　　　　　　　
R. Kuriyama; M. Imashimizu; M. Kuramochi
第46回日本分子生物学会年会, Dec. 2023

生きた線虫C. elegans細胞内における蛍光タンパク質結晶　　　　　　　　　　　　　　　
I. Sugawara; Y. Shinkai; M. Kuramochi
第46回日本分子生物学会年会, Dec. 2023

X線回折で見る高分子材料内部フィラーの回転ダイナミクス　　　　　　　　　　　　　　　
新井 達也; 倉持 昌弘; 三尾 和弘; 関口 博史; 佐々木 裕次
第７２回高分子討論会, Sep. 2023

Functional analysis of the ice-binding proteins and their application to bio-preservation technology　　　　　　　　　　　　　　　
M. Kuramochi
Tsukuba conference 2023, Sep. 2023, [Invited]

Ice-Binding Proteins: Structure-specific functions in vivo　　　　　　　　　　　　　　　
M. Kuramochi
9IDMRCS, Aug. 2023, [Invited]

Dynamical observation of the crystallized protein in the living Caenorhabditis elegans cells　　　　　　　　　　　　　　　
Ibuki Sugawara; Yoichi Shinkai; Masahiro Kuramochi
9IDMRCS, Aug. 2023

Investigation of non-thermal effects of sub-terahertz on living Caenorhabditis elegans　　　　　　　　　　　　　　　
Ryohei Kuriyama; Masahiko Imashimizu; Masahiro Kuramochi
9IDMRCS, Aug. 2023

凍結低温制御分子の構造特異機能の解明および個体丸ごと保存技術の開発　　　　　　　　　　　　　　　
倉持昌弘
ACT-X「環境とバイオテクノロジー」第5回領域会議, Aug. 2023

氷晶結合タンパク質を用いた基礎応用研究　　　　　　　　　　　　　　　
倉持昌弘
TRiSTARイベント 「茨大から世界へ―分野・業種・国境... あらゆるボーダーを越えていくために」, Apr. 2023

機械学習を用いたX線動画解析による動態物性の抽出　　　　　　　　　　　　　　　
倉持昌弘; 李卓起; 赤穂昭太郎; 佐々木裕次
2022年度量子ビームサイエンスフェスタ, Mar. 2023

凍結低温制御分子の構造特異機能の解明および個体丸ごと保存技術の開発　　　　　　　　　　　　　　　
倉持昌弘
ACT-X「環境とバイオテクノロジー」第４回領域会議, Jan. 2023

Twisting Motions of Different Ligand-bound States of nAChR α7 Monitored by Diffracted X-ray Tracking　　　　　　　　　　　　　　　
楊 越; 新井 達也; 佐々木 大; 倉持 昌弘; 関口 博史; 三尾 和弘; 久保 泰; 佐々木 裕
第３５回日本放射光学会年会, Jan. 2023

不凍タンパク質を用いた研究展開 細胞、生体組織、個体生物の保存技術へ　　　　　　　　　　　　　　　
倉持昌弘
TRiSTARトランスボーダーサイエンスキャンプ, Jan. 2023

類似分子からなるキラル結晶とラセミ結晶の結晶構造と力学特性の比較　　　　　　　　　　　　　　　
石崎一輝; 高木大輔; 朝日透; 倉持昌弘; 谷口卓也
日本結晶学会年会講演要旨集, 2023
2023, 2023

細胞内液-液相分離の攪乱とその生体応答　　　　　　　　　　　　　　　
丸山迪代; 三村真大; 三村真大; 新木和孝; 井上明俊; 倉持昌弘; 戸井基道; 草田裕之; 冨田峻介; 新海陽一
日本生化学会大会(Web), 2023
2023, 2023

"氷結合タンパク質を発現した線虫の凍結保存効果 Cryopreservation effects of C. elegans expressing ice-binding proteins"　　　　　　　　　　　　　　　
下谷祐平; 新井達也; 三尾和弘; 津田栄; 佐々木裕次; 倉持昌弘
第45回日本分子生物学会年会, Dec. 2022

X線ブリンキング法による動態物性取得と機能相関 生体内分子、高分子、ソフトクリスタルまで　　　　　　　　　　　　　　　
倉持昌弘; 三尾和弘; 佐々木裕次
2022年度OPERANDO-OIL・COMS・量子ビーム計測クラブ合同研究会, Nov. 2022, [Invited]

Dynamical Measurement of Material Characteristics using Diffracted X-ray Blinking　　　　　　　　　　　　　　　
M. Kuramochi
ICPAC KK2022, Nov. 2022, [Invited]

X 線 1 分子追跡法によるイベルメクチン存在下での nAChR α7 の逆回転運動の測定　　　　　　　　　　　　　　　
Yue Yang; Tatsuya Arai; Daisuke Sasaki; Masahiro Kuramochi; Hiroshi Sekiguchi; Kazuhiro Mio; Tai Kubo; Yuji C. Sasaki
第６０回日本生物物理学会, Sep. 2022

Mechanism of in-cell protein crystallization with supersaturated environment and its application　　　　　　　　　　　　　　　
倉持昌弘
分子夾雑の生命化学 成果とりまとめ公開シンポジウム, Sep. 2022

不凍タンパク質の生体内機能解析およびX線ブリンキングによる分子動態計測　　　　　　　　　　　　　　　
倉持昌弘
TRiSTAR 育成支援対象者間の交流の場 ・ 異分野クロスオーバー, Sep. 2022

味覚神経ASE を介したAIY 神経の活動制御　　　　　　　　　　　　　　　
倉持昌弘; 山口一輝
線虫研究の未来を創る会2022, Aug. 2022

サンゴ由来蛍光結晶化遺伝子Xpa 導入による線虫体内での結晶形成　　　　　　　　　　　　　　　
菅原生蕗; 倉持昌弘
線虫研究の未来を創る会2022, Aug. 2022

テラヘルツ光照射による線虫初期胚の非熱効果検証　　　　　　　　　　　　　　　
栗山稜平; 倉持昌弘
線虫研究の未来を創る会2022, Aug. 2022

不凍タンパク質の凍結保存効果と水和構造に起因した氷結合メカニズム　　　　　　　　　　　　　　　
下谷祐平; 倉持昌弘
線虫研究の未来を創る会2022, Aug. 2022

Expression of Ice-Binding Proteins in Caenorhabditis elegans Improves the Survival Rate after freezing and cold shock.　　　　　　　　　　　　　　　
M. Kuramochi
IBP conference, Aug. 2022

"Functional analysis in vivo of ice-binding proteins based on their dynamic behaviour. 氷結合タンパク質の動的挙動に基づいた生体内機能解析"　　　　　　　　　　　　　　　
倉持 昌弘
日本蛋白質学会PSSJ2022, Jun. 2022

Cryopreservation effects of C. elegans expressing ice-binding proteins　　　　　　　　　　　　　　　
下谷祐平; 新井達也; 三尾和弘; 津田栄; 佐々木裕次; 倉持昌弘
日本分子生物学会年会プログラム・要旨集(Web), 2022
2022, 2022

時分割回折X線ブリンキングによるソフトクリスタル動的特性観測　　　　　　　　　　　　　　　
倉持 昌弘
新学術領域研究「ソフトクリスタル」第8回公開シンポジウム, 2022

回折X線ブリンキングによる結晶材料の動的構造解析　　　　　　　　　　　　　　　
倉持昌弘、引田理英、佐々木裕次
2021年度量子ビームサイエンスフェスタ, 2022

線虫C. エレガンスにおける凝集タンパク質および分子夾雑場のX線1分子観察　　　　　　　　　　　　　　　
倉持 昌弘
新学術領域研究「分子夾雑化学」第6回領域会議, 2022

光による結晶のねじれ発生と動的挙動解析　　　　　　　　　　　　　　　
谷口卓也; 石崎一輝; 高木大輔; 重宗宏毅; 西村一紀; 倉持昌弘; 佐々木裕次; 小島秀子; 朝日透; 朝日透
日本結晶学会年会講演要旨集, 2021
2021, 2021

X線ブリンキング法による⽣物個体内の結晶化タンパク質および分⼦夾雑場観察　　　　　　　　　　　　　　　
倉持 昌弘
3新学術領域合同シンポジウム, 2021

Cryopreservation of C. elegans by ice-binding protein　　　　　　　　　　　　　　　
下谷祐平、倉持昌弘
線虫研究の未来を創る会, 2021

X-ray avoidance behaviour in C. elegans　　　　　　　　　　　　　　　
大津乃亜、倉持昌弘
線虫研究の未来を創る会, 2021

Mechanism of in-cell protein crystallization with supersaturated environment and its application　　　　　　　　　　　　　　　
倉持 昌弘
線虫研究の未来を創る会, 2021

時分割回折X線ブリンキングによるソフトクリスタル動的特性観測　　　　　　　　　　　　　　　
倉持 昌弘
新学術領域研究「ソフトクリスタル」第7回公開シンポジウム, 2021

線虫C. エレガンスにおける凝集タンパク質および分子夾雑場のX線1分子観察　　　　　　　　　　　　　　　
倉持 昌弘
新学術領域研究「分子夾雑化学」第5回領域会議, 2021

固体高分子結晶性電解質内部のオペランド分子動態計測 Operand-Measurement of molecular dynamics inside Solid Polymer Crystalline Electrolytes　　　　　　　　　　　　　　　
佐々木大輔、橘川莉子、中山美沙恵、倉持昌弘、一柳光平、深谷亮、関口博史、佐藤文菜、野澤俊介、足立伸一、三尾和弘、 田所誠、佐々木裕次
2020年度 量子ビームサイエンスフェスタ, 2021

DXB 法を用いた不凍タンパク質結晶の内部分子動態計測 Molecular motion of antifreeze protein in crystal measured by Diffracted X-ray Blinking　　　　　　　　　　　　　　　
新井達也、倉持昌弘、三尾和弘、佐々木裕次
2020年度 量子ビームサイエンスフェスタ, 2021

結晶性/非晶性 PEEK 高分子膜の時分割動態計測 Time-resolved observations of crystal / non-crystal PEEK film　　　　　　　　　　　　　　　
下村峻矢、倉持昌弘、佐々木大輔、新井達也、 引田理英、佐々木裕次
2020年度 量子ビームサイエンスフェスタ, 2021

回折X線ブリンキングによるタイヤゴムの動的構造解析 Dynamic Structure Analysis for Tire Rubbers by Diffracted X-ray Blinking　　　　　　　　　　　　　　　
倉持昌弘、増井友美、間下亮、引田理英、新井達也、 三尾和弘、関口博史、岸本浩通、佐々木裕次
2020年度 量子ビームサイエンスフェスタ, 2021

生きた細胞上の GPCR のミリ秒-マイクロ秒ダイナミクスMilli- to microsecond dynamics of Single GPCR in Living Cells　　　　　　　　　　　　　　　
藤村章子、石原正輝、一柳光平、野澤俊介、足立伸一、深谷亮、倉持昌弘、関口博史、三尾和弘、佐々木裕次
2020年度 量子ビームサイエンスフェスタ, 2021

時分割回折X線ブリンキングによるソフトクリスタル動的特性観測　　　　　　　　　　　　　　　
M Kuramochi
新学術領域研究「ソフトクリスタル」共同研究推進会議 2, 2020

線虫C. エレガンスにおける凝集タンパク質および分子夾雑場のX線1分子観察　　　　　　　　　　　　　　　
M Kuramochi
新学術領域研究「分子夾雑化学」第8回オンラインセミナー, 2020

In vivo X-ray single-molecule observation of ice-binding proteins for Caenorhabditis elegans freezing tolerance　　　　　　　　　　　　　　　
M Kuramochi
第20回日本蛋白質科学会年会, 2020

時分割回折X線ブリンキングによるソフトクリスタル動的特性観測　　　　　　　　　　　　　　　
M Kuramochi
新学術領域研究「ソフトクリスタル」第７回領域全体会議, 2020

ROTATIONAL MOTION OF SINGLE TRPV1 CHANNEL UPON GATING　　　　　　　　　　　　　　　
Shoko Fujimura, Kazuhiro Mio, Masahiro Kuramochi, Hiroshi Sekiguchi, Muneyo Mio, Tai Kubo, Yuji C. Sasaki
米国生物物理学会, 2020

DYNAMIC INTERNAL MOTION OF GPCR ON LIVE CELLS　　　　　　　　　　　　　　　
Masaki Ishihara, Shoko Fujimura, Kohei Ichiyanagi, Shunsuke Nozawa, Shinichi Adachi, Ryo Fukaya, Masahiro Kuramochi, Hiroshi Sekiguchi, Kazuhiro Mio, Yuji C. Sasaki
米国生物物理学会, 2020

DIRECT DETECTION OF INTRAMOLECULAR DYNAMICS OF MEMBRANE PROTEINS USING TIME-RESOLVED X-RAY SINGLE-MOLECULE TRACKING　　　　　　　　　　　　　　　
Kazuhiro Mio1, Shoko Fujimura1, Masaki Ishihara2, Muneyo Mio1, Masahiro Kuramochi2, Hiroshi Sekiguchi3, Tai Kubo1, Yuji C. Sasaki2.
米国生物物理学会, 2020

PROTEIN CRYSTAL MOTIONS FROM TIME-RESOLVED DIFFRACTED X-RAY BLINKING　　　　　　　　　　　　　　　
Yuji C. Sasaki, Masahiro Kuramochi, Kazuhiro Mio, Hiroshi Sekiguchi, Ayana Sato-Tomita, Naoya Shibayama
米国生物物理学会, 2020

SINGLE MOLECULAR OBSERVATION OF AFP AND ICE-CRYSTAL DYNAMICS IN CAENORHABDITIS ELEGANS BY TIME-RESOLVED X-RAY DIFFRACTION MEASUREMENTS　　　　　　　　　　　　　　　
Yige Dong, Masahiro Kuramochi, Chiaki Takanashi, Kazuhiro Mio, Motomichi Doi, Kouki Aoyama, Hiroshi Sekiguchi, Sakae Tsuda, Yuji C. Sasaki
米国生物物理学会, 2020

THE OBSERVATION OF THE ICE-BINDING PROTEIN FOR FREEZING AND COLD TOLERANCE OF TRANSGENIC CAENORHABDITIS ELEGANS　　　　　　　　　　　　　　　
Masahiro Kuramochi, Yoichi Shinkai, Motomichi Doi, Kazuhiro Mio, Sakae Tsuda, Yuji C. Sasaki
米国生物物理学会, 2020

回折Ｘ線明滅法（DXB）を用いた歯車型分子結晶からの分子動態測定　　　　　　　　　　　　　　　
佐々木大輔 、佐々木裕次、倉持昌弘、石原正輝、尾股紘希、田所誠、亀渕萌、菅大樹
第33回日本放射光学会年会・放射光科学合同シンポジウム, 2020

生細胞上 GPCR1 分子の時間依存的内部運動計測　　　　　　　　　　　　　　　
石原正輝、藤村章子、一柳光平、野澤俊介、足立伸一 、深谷亮、倉持昌弘、関口博史、 三尾和弘、佐々木裕次
第33回日本放射光学会年会・放射光科学合同シンポジウム, 2020

時分割回折Ｘ線法による線虫Ｃ．エレガンスの不凍タンパク質と氷の１分子動態観察　　　　　　　　　　　　　　　
董芸格、倉持昌弘、高梨千晶、三尾和弘、戸井基道、青山光輝、関口博史、津田栄、佐々木裕次
第33回日本放射光学会年会・放射光科学合同シンポジウム, 2020

生きた線虫体内の不凍タンパク質動態を捉える生体内時分割１分子測定　　　　　　　　　　　　　　　
倉持昌弘; 高梨千晶; 関口博史; 青山光輝; 戸井基道; 三尾和弘; 津田栄; 佐々木裕次
第32回日本放射光学会年会・放射光科学合同シンポジウム, 2019

明滅する回折Ｘ線を用いた結晶動態計測　　　　　　　　　　　　　　　
尾股; 水牧; 河村; 大沢; 鈴木; 関口; 倉持; 三尾; 佐々木
第32回日本放射光学会年会・放射光科学合同シンポジウム, 2019

生きた細胞上のGPCR１分子動態計測　　　　　　　　　　　　　　　
石原; 藤村; 一柳; 野澤; 足立; 深谷; 倉持; 三尾; 佐々木
第32回日本放射光学会年会・放射光科学合同シンポジウム, 2019

TRPV1チャネルの１分子内回転動態の決定　　　　　　　　　　　　　　　
藤村 章子; 三尾 和弘; 倉持 昌弘; 関口 博史; 三尾 宗代; 久保 泰; 佐々木 裕次
第32回日本放射光学会年会・放射光科学合同シンポジウム, 2019

Detecting Intramolecular Dynamics of GPCRs in Living Cells Using Diffracted X-ray Blinking Technique　　　　　　　　　　　　　　　
Masaki ISHIHARA, Shoko FUJIMURA, Kouhei ICHIYANAGI, Shunsuke NOZAWA, Shinichi ADACHI, Ryo FUKAYA,  Masahiro KURAMOCHI, Kazuhiro MIO, Yuji C SASAKI
MATERIALS RESEARCHMEETING 2019, 2019

Nanocrystal Dynamics using Time-resolved Diffracted X-ray Blinking　　　　　　　　　　　　　　　
YUJI SASAKI, M. KURAMOCHI, H. SEKIGUCHI
MATERIALS RESEARCHMEETING 2019, 2019

Dynamics And Functions of Antifreeze Proteins In Transgenic Caenorhabditis Elegans At Freezing Environments　　　　　　　　　　　　　　　
M. Kuramochi, G. Tou, C. Takanashi, A. Yamauchi, M. Doi, K. Mio, S. Tsuda, H. Sekiguchi, Y. C. Sasaki
ASCB-EMBO2019, 2019

MHCIIのユビキチン化は運動性の変化により抗原提示の傾向を変える　　　　　　　　　　　　　　　
河本 隆志1、Yao Yilun1、小園 裕子1、倉持 昌弘2、佐々木 裕次2、小園 晴生
第４２回日本分子生物学会年会, 2019

時分割回折Ｘ線計測による線虫Ｃ．エレガンスの不凍タンパク質1分子動態観察　　　　　　　　　　　　　　　
董 芸格1、倉持 昌弘1,2,4、高梨 千晶1、三尾 和弘2、戸井 基道4、青山 光輝3、関口 博史3、津田 栄4、佐々木 裕次
第４２回日本分子生物学会年会, 2019

X線1分子追跡法によるTRPV1チャネル回転動態の決定　　　　　　　　　　　　　　　
三尾 和弘1、藤村 章子1、倉持 昌弘2、関口 博史3、三尾 宗代1、久保 泰1、佐々木 裕次
第４２回日本分子生物学会年会, 2019

X線1分子計測法を用いた生細胞上のGPCR時間依存的分子内部運動の検出　　　　　　　　　　　　　　　
石原 正輝1,2、藤村 章子2、一柳 光平3,4、野澤 俊介3、足立 伸一3、深谷 亮3、倉持 昌弘1,2、三尾 和弘2、佐々木 裕
第４２回日本分子生物学会年会, 2019

氷結合タンパク質変異導入による線虫Ｃ．エレガンスの低温耐性への影響 The in vivo effects of the ice-binding protein mutations for the cold tolerance in C. elegans　　　　　　　　　　　　　　　
倉持 昌弘、董 芸格、高梨 千晶、山内 彩加林、戸井 基道、三尾 和弘、津田 栄、佐々木 裕次
第４２回日本分子生物学会年会, 2019

Individual single nanocrystal motions observed using time-resolved diffracted x-ray blinking　　　　　　　　　　　　　　　
Hiroki Omata, Masahiro Kuramochi, Hiroshi Sekiguchi and Yuji C Sasaki
12th International Symposium on Atomic Level Characterizations for New Materials and Devices ‘19, 2019

膜タンパク質の分子内部動態解析技術の開発 Understanding intramolecular dynamics of membrane proteins using X-ray based analysis techniques　　　　　　　　　　　　　　　
Kazuhiro Mio, Shoko Fujimura, Masaki Ishihara, Muneyo Mio, Masahiro Kuramochi, Hiroshi Sekiguchi, Tai Kubo, Yuji C. Sasaki
第57回日本生物物理学会年会, 2019

TRPV1 分子内部の回転動態の決定 Agonist- and antagonist-induced rotational motion of TRPV1 channel　　　　　　　　　　　　　　　
Shoko Fujimura, Kazuhiro Mio, Masahiro Kuramochi, Sekiguchi Hiroshi, Muneyo Mio, Tai Kubo, Yuji Sasaki
第57回日本生物物理学会年会, 2019

回折 X 線ブリンキング法を用いた生細胞上の GPCR 分子内部運動の決定 Determining Intramolecular Motion of GPCRs on Live Cells using Diffracted X-ray Blinking Technique　　　　　　　　　　　　　　　
Masaki Ishihara, Shoko Fujimiura , Kohei Ichiyanagi, Shunsuke Nozawa, Shinichi Adachi, Ryo Fukaya, Masahiro Kuramochi, Kazuhiro Mio, Yuji Sasaki
第57回日本生物物理学会年会, 2019

非凍結温度において氷結合タンパク質は線虫の低温耐性を改善する Ice-Binding Proteins Improves the Survival Rate of Caenorhabditis elegans at Non-freezing Temperature　　　　　　　　　　　　　　　
Masahiro Kuramochi, Geikaku Tou, Chiaki Takanashi, Motomichi Doi, Kazuhiro Mio, Sakae Tsuda, C. Yuji Sasaki
第57回日本生物物理学会年会, 2019

Structure, Dynamics, Function and in vivo effects of Antifreeze protein　　　　　　　　　　　　　　　
倉持昌弘
線虫研究の未来を創る会 2019, 2019

生きた細胞上の GPCR1 分子 X 線動態計測 X-ray single molecular observation of GPCRs in living cells　　　　　　　　　　　　　　　
石原 正輝、藤村 章子、一柳 光平、野澤 俊介、足立 伸一、深谷 亮、倉持 昌弘、三尾 和弘、佐々木 裕次
第19回日本蛋白質科学会年会, 2019

TRPV1 チャネルの 1 分子内回転動態の決定 Rotational Motions of Single TRPV1 Channel upon Gating　　　　　　　　　　　　　　　
三尾 和弘、藤村 章子、倉持 昌弘、関口 博史、三尾 宗代、久保 泰、佐々木 裕次
第19回日本蛋白質科学会年会, 2019

凍結・非凍結環境における氷結合タンパク質の生体内作用機序解明にむけて 線虫Ｃ．エレガンスの低温耐性観察およびＸ線１分子動態測定 In vivo effect of Ice-Binding Protein Cold tolerance observation and X-ray single molecular measurement using living C. elegans　　　　　　　　　　　　　　　
倉持 昌弘、董 芸格、高梨 千晶、山内 彩加林、戸井 基道、青山 光輝、関口 博史、三尾 和弘、津田 栄、佐々木 裕次
第19回日本蛋白質科学会年会, 2019

神経個性を決める潜在的クロマチン変化の意義とその制御機構の解明　　　　　　　　　　　　　　　
新海 陽一、倉持 昌弘、戸井 基道
新学術領域研究「クロマチン潜在能」第二回領域会議, 2019

前もって準備されたクロマチン変化は神経細胞の成熟や個性決定に重要である-in vivo単一遺伝子座ヒストン修飾可視化法を用いた解析-　　　　　　　　　　　　　　　
新海 陽一、倉持 昌弘、戸井 基道
第13回日本エピジェネティクス研究会年会, 2019

Single molecular measurements of GPCRs in living cells　　　　　　　　　　　　　　　
2019

Natural Killer 細胞を用いたＸ線 1 分子動態観察 X-ray Single Molecule Observations of Natural Killer Cell　　　　　　　　　　　　　　　
張宰源 1, 2、Y. Baek3 , 倉持昌弘 1, 2、関口博史 4、一柳光平 5、池崎圭吾 6、李仁 志 3、佐々木裕次 1
量子ビームサイエンスフェスタ, 2019

POINT MUTATION OF THE ICE-BINDING SITE IN ANTIFREEZE PROTEIN MODIFY THE COLD TOLERANCE IN CAENORHABDITIS ELEGANS　　　　　　　　　　　　　　　
Masahiro Kuramochi, Chiaki Takanashi, Akari Yamauchi, Motomichi Doi, Kazuhiro Mio, Sakae Tsuda, Yuji C. Sasaki
63rd annual meeting  Biophysical Society (BPS19), 2019

DIFFRACTED X-RAY BLINKING FROM NANOCRYSTAL ON PROTEIN USED AS INTERNAL MOTION PROBE　　　　　　　　　　　　　　　
Hiroshi Sekiguchi, Masahiro Kuramochi, Yuji C. Sasaki
63rd annual meeting  Biophysical Society (BPS19), 2019

MICRO-SECOND X-RAY SINGLE MOLECULE DYNAMICS OF ALLOSTERIC TWISTING MOTIONS IN HEMOGLOBIN　　　　　　　　　　　　　　　
Yuji C. Sasaki, Masahiro Kuramochi, Yuu Okamura, Hiroshi Sekiguchi, Naoki Yamamoto, Naoya Shibayama
63rd annual meeting  Biophysical Society (BPS19), 2019

ANTAGONIST-INDUCED CLOCKWISE ROTATION IN THE TRPV1　　　　　　　　　　　　　　　
Shoko Fujimura, Kazuhiro Mio, Masahiro Kuramochi, Hiroshi Sekiguchi, Muneyo Mio, Tai Kubo, Yuji C. Sasaki
63rd annual meeting  Biophysical Society (BPS19), 2019

DETECTING INTRAMOLECULAR DYNAMICS OF GPCRS USING DIFFRACTED X-RAY BLINKING TECHNIQUE　　　　　　　　　　　　　　　
Kazuhiro Mio, Masaki Ishihara, Shoko Fujimura, Masahiro Kuramochi, Yuji C. Sasaki
63rd annual meeting  Biophysical Society (BPS19), 2019

UBIQUITINATION OF MHC II CHANGES DYNAMICS OF ITS RECOGNITION STRUCTURE　　　　　　　　　　　　　　　
Haruo Kozono, Takashi Kawamoto, Yuko Kozono, masahiro kuramochi, Yuji C. Sasaki
63rd annual meeting  Biophysical Society (BPS19), 2019

X線1分子追跡法によるTRPV1チャネル回転動態の決定　　　　　　　　　　　　　　　
三尾 和弘; 藤村 章子; 倉持 昌弘; 関口 博史; 三尾 宗代; 久保 泰; 佐々木 裕次
第41回日本分子生物学会年会, 2018

時分割X線回折ブリンキングによる生体内１分子動態の観察　　　　　　　　　　　　　　　
倉持 昌弘; 石原 正輝; 藤村 章子; 戸井 基道; 三尾 和弘; 佐々木 裕次
第41回日本分子生物学会年会, 2018

不凍タンパク質は低温環境下における線虫の細胞を保護し低温耐性を強化する　　　　　　　　　　　　　　　
高梨千晶; 倉持昌弘; 山内彩加林; 戸井基道; 三尾和弘; 津田栄; 佐々木裕次
第41回日本分子生物学会年会, 2018

生きた細胞上のGPCR１分子動態計測に成功　　　　　　　　　　　　　　　
石原; 藤村; 倉持; 三尾; 佐々木
新学術領域研究「３D活性サイト科学」第8回成果報告会, 2018

不凍分子活性サイトを改変して動物の低温耐性を制御する　　　　　　　　　　　　　　　
倉持; 高梨; 山内; 戸井; 三尾; 津田; 佐々木
新学術領域研究「３D活性サイト科学」第8回成果報告会, 2018

Ubiquitination of MHCII changes tendency of antigen presentation due to structral conversion of MHCII　　　　　　　　　　　　　　　
河本 隆志; 小園 裕子; 張 宰源; 倉持 昌弘; 佐々木 裕次; 小園 晴生
日本生物物理学会年会, 2018

3D motion of TRPV1 cation channel depicted by diffracted X-ray tracking method X線1分子追跡法によるTRPV1チャネルの3次元運動　　　　　　　　　　　　　　　
藤村 章子; 三尾 和弘; 倉持 昌弘; 関口 博史; 三尾 宗代; 久保 泰; 佐々木 裕次
日本生物物理学会年会, 2018

Protein Motion Analyzed by Diffracted X-ray Blinking　　　　　　　　　　　　　　　
関口 博史; 倉持 昌弘; 太田 昇; 佐々木 裕次
日本生物物理学会年会, 2018

High-speed single molecule tracking of allosteric transitions in hemoglobin using Diffracted X-ray Tracking (DXT)　　　　　　　　　　　　　　　
岡村 優; 倉持 昌弘; 開 俊樹; 山本 直樹; 柴山 修哉; 関口 博史; 佐々木 裕次
日本生物物理学会年会, 2018

The ice-binding proteins protect the cells and contribute to increase the survival rate in Caenorhabditis elegans under cold environments 不凍タンパク質は低温環境下における線虫の細胞を保護し、生存率を上昇させる　　　　　　　　　　　　　　　
倉持 昌弘; 高梨 千晶; 山内 彩加林; 津田 栄; 戸井 基道; 三尾 和弘; 佐々木 裕次
第56回日本生物物理学会年会, 2018

X線1分子追跡法による生体内分子動態の観察　　　　　　　　　　　　　　　
倉持昌弘
「線虫研究の未来を創る会」, 2018

カルシウムイメージングによる線虫神経回路網の動態解析　　　　　　　　　　　　　　　
戸井 基道; 倉持 昌弘
計測自動制御学会 ライフエンジニアリング部門シンポジウム2018（2B4 脳神経回路の動作を見て測る）, 2018

X線１分子追跡法によるTRPV１チャネルの３次元運動　　　　　　　　　　　　　　　
藤村章子; 三尾和弘; 倉持昌弘; 関口博史; 三尾宗代; 久保 泰; 佐々木裕次
バイオイメージング学会, 2018

X線１分子追跡法による線虫１分子動態の観察　　　　　　　　　　　　　　　
倉持昌弘; 関口博史; 青山光輝; 戸井基道; 三尾和弘; 津田栄; 佐々木裕次
第27回バイオイメージング学会, 2018

ヘモグロビン・アロステリック 1 分子転移計測 Single Molecular Measurements on the Allosteric Transitions of Hemoglobin　　　　　　　　　　　　　　　
岡村優; 倉持昌弘; 開俊樹; 柴山修哉; 関口博史; 佐々木裕次
日本タンパク質学会年会, 2018

不凍タンパク質は凍結・低温環境下における線虫 C．エレガンスの生存率を上昇させる Antifreeze protein incerase the survival rate of C. elegans in freeze environments and low temperatures　　　　　　　　　　　　　　　
倉持昌弘; 高梨千晶; 戸井基道; 関口博史; 津田栄; 佐々木裕次
第18回日本タンパク質学会年会, 2018

線虫C. elegansを用いた不凍タンパク質のin vivo解析　　　　　　　　　　　　　　　
倉持昌弘; 高梨千晶; 山内彩加林; 戸井基道; 三尾和弘; 津田栄; 佐々木裕次
低温生物工学会, 2018

Ｘ線1分子追跡法とX線輝点明滅法によるTRPV1の作動機構解析　　　　　　　　　　　　　　　
藤村章子; 三尾和弘; 倉持昌弘; 池崎圭吾; 松原 賢; 関口博史; 久保 泰; 佐々木裕次
生理学研究所 研究会 TRPチャネル～免疫・炎症応答の重要な媒介分子～, 2018

タンパク質・分子内運動計測に向けたX線１分子追跡法の低侵襲化および角度ダイナミックレンジ広範化　　　　　　　　　　　　　　　
関口博史; 青山光輝; 倉持昌弘; 池崎圭吾; 佐々木裕次
新学術領域研究「３D活性サイト科学」第７回成果報告会, 2018

線虫C. elegansを用いた不凍タンパク質の細胞機能解析、および生体内Ｘ線１分子追跡法によるタンパク質動態の観察　　　　　　　　　　　　　　　
倉持昌弘; 高梨千晶; 関口博史; 戸井基道; 津田栄; 佐々木裕次
新学術領域研究「３D活性サイト科学」第７回成果報告会, 2018

MICROSCOPIC OBSERVATIONS OF PROTEIN BROWNIAN MOTIONS IN SUPERSATURATED SOLUTIONS　　　　　　　　　　　　　　　
Kazuki Yoshimura; Masahiro Kuramochi; Yuji C. Sasaki
62nd annual meeting Biophysical Society (BPS18), 2018

IN VIVO X-RAY MONITORING OF DYNAMICS BETWEEN INTERLEUKIN 2 AND INTERLEUKIN 15 ON NK CELLS　　　　　　　　　　　　　　　
Jaewon Chang; Masahiro Kuramochi; Youngsuk Beak; Kouhei Ichiyanagi; Hiroshi Sekiguchi; Yuji C. Sasaki
62nd annual meeting Biophysical Society (BPS18), 2018

FUNCTIONAL ANALYSIS OF ANTIFREEZE PROTEINS FOR COLD TOLERANCE BEHAVIOR and X-ray single molecule observations IN C. ELEGANS　　　　　　　　　　　　　　　
M. Kuramochi; C. Takanashi; H. Sekiguchi; M. Doi; S. Tsuda; Y. C. Sasaki
62nd annual meeting Biophysical Society (BPS18), 2018

ROTATIONAL BROWNIAN MOTION OF TRPV1 CHANNEL OBSERVED BY SYNCHROTRON DIFFRACTED X-RAY TRACKING AND LABORATORY X-RAY BLINKING ANALYSIS　　　　　　　　　　　　　　　
Kazuhiro Mio; Masahiro Kuramochi; Ken Matsubara; Keigo Ikezaki; Muneyo Mio; Hiroshi Sekiguchi; Tai Kubo; Yuji C. Sasaki
62nd annual meeting Biophysical Society (BPS18), 2018

NOVEL IN VIVO OBSERVATIONS OF SINGLE PROTEIN MOTIONS USING LABORATORY X-RAY SOURCE　　　　　　　　　　　　　　　
Yuji C. Sasaki; M Kuramochi; H. Sekiguchi; K. Mio
62nd annual meeting Biophysical Society (BPS18), 2018

生体内における不凍タンパク質動態のX線1分子追跡　　　　　　　　　　　　　　　
高梨千晶; 倉持昌弘; 関口博史; 一柳光平; 佐々木裕次
第３１回日本放射光学会年会, 2018

NK細胞膜上IL-2とIL-15のin vivo X線運動計測　　　　　　　　　　　　　　　
張宰源; 倉持昌弘; 一柳光平; 関口博史; 佐々木裕次
第３１回日本放射光学会年会, 2018

単色X線によるTRPV1のカプサイシン分子内応答運動解析　　　　　　　　　　　　　　　
松原賢; 倉持昌弘; 関口博史; 一柳光平; 三尾和弘; 佐々木裕次
第３１回日本放射光学会年会, 2018

ヘモグロビン・アロステリック 1 分子転移計測　　　　　　　　　　　　　　　
岡村優; 倉持昌弘; 開俊樹; 柴山修哉; 関口博史; 佐々木裕次
第３１回日本放射光学会年会, 2018

X線ブリンキングによるタンパク質1分子動態の解析　　　　　　　　　　　　　　　
倉持昌弘; 関口博史; 佐々木裕次
第３１回日本放射光学会年会, 2018

線虫の低温耐性から見た不凍タンパク質の機能解析、および生体内X線１分子追跡法によるAFP分子動態の解析 Functional analysis of antifreeze protein (AFP) for cold tolerance in C. elegans, and the analysis of AFP molecular dynamics by in vivo diffracted X-ray tracking　　　　　　　　　　　　　　　
M. Kuramochi; C. Takanashi; H. Sekiguchi; M. Doi; S. Tsuda; Y. C. Sasaki
2017年度生命科学系学会合同年次大会, 2017

X 線 1 分子追跡法による TRPV1 チャネルの分子運動解析 3D MOTION MAPS OF TRPV1 CATION CHANNEL DEPICTED BY DIFFRACTED X-RAY TRACKING METHOD　　　　　　　　　　　　　　　
Kazuhiro Mio; Keigo Ikezaki; Masahiro Kuramochi; Hiroshi Sekiguchi; Tai Kubo; Yuji C. Sasaki
第５５回日本生物物理学会年会, 2017

生自然殺害細胞内 X 線 1 分子計測 X-ray single molecular observations in living Natural killer cells　　　　　　　　　　　　　　　
張 宰源; 倉持 昌弘; 一柳 光平; 佐々木 裕次
第５５回日本生物物理学会年会, 2017

In vivo X-ray single molecule observation and functional analysis of antifreeze proteins for cold tolerance in C. elegans 線虫C. elegansの低温耐性から見た不凍タンパク質の機能解析、および生体内X線一分子観察　　　　　　　　　　　　　　　
倉持 昌弘; 高梨 千晶; 関口 博史; 戸井 基道; 津田 栄; 佐々木 裕次
第５５回日本生物物理学会年会, 2017

X線1分子追跡法 (Diffracted X-ray Tracking)を用いたTRPV1のカプサイシン応答分子運動解析　　　　　　　　　　　　　　　
三尾和弘; 倉持昌弘; 池崎圭吾; 関口博史; 久保泰; 佐々木 裕次
生理学研究所 研究会, TRPチャネル～オルガネラ Ca2+シグナルの重要な媒介分子～, 2017

Excitatory/inhibitory switch from asymmetric sensory neurons defines postsynaptic tuning for rapid response in the C. elegans salt-chemotaxis circuit 線虫の非対称機能をもつ味覚神経からの興奮性/抑制性シナプス伝達が塩走化性を制御する介在神経の迅速な応答を調節する　　　　　　　　　　　　　　　
倉持昌弘; 戸井基道
第４０回日本神経科学大会, 2017

Excitatory/inhibitory switch from asymmetric sensory neurons defines postsynaptic tuning for rapid response in the C. elegans salt-chemotaxis circuit.　　　　　　　　　　　　　　　
Masahiro Kuramochi; Motomichi Doi
21st International C. elegans Meeting, 2017

味覚神経ASEからの興奮性/抑制性シナプス伝達が介在神経AIBの迅速な応答を調節する　　　　　　　　　　　　　　　
倉持昌弘; 戸井基道
東京地区線虫勉強会, 2017

An integrated analysis for the dynamical responses of pre- and postsynaptic neurons through synaptic encoding in the C. elegans salt-chemotaxis circuit　　　　　　　　　　　　　　　
M. Kuramochi; M. Doi
Society for Neuroscience (SfN) 2016, Nov. 2016

A quantitative description for the dynamical responses of postsynaptic neurons through synaptic encoding in the C. elegans salt-chemotaxis circuit　　　　　　　　　　　　　　　
M. Kuramochi; M. Doi
CeNeuro2016, Jul. 2016

カルシウムイメージングと計算機シミュレーションによる線虫の塩化走性に関わる神経コーディングの時空間ダイナミクス　　　　　　　　　　　　　　　
倉持 昌弘; 戸井 基道
第38回日本神経科学大会, Jul. 2015

Calcium imaging and computational simulation for the spatio-temporal dynamics of the neural coding in a C. elegans chemotaxis circuit　　　　　　　　　　　　　　　
M. Kuramochi; M. Doi
20th International C. elegans Meeting, Jun. 2015

線虫C. elegansの味覚神経活動の時空間動態解析　　　　　　　　　　　　　　　
倉持 昌弘; 戸井 基道
電子情報通信学会 ニューロコンピューティング研究会, Mar. 2015

Integrated analysis of spatio-temporal dynamics in a C. elegans chemosensory circuit　　　　　　　　　　　　　　　
倉持 昌弘; 戸井 基道
第３７回日本分子生物学会年会, Nov. 2014

Calcium imaging, dynamical modeling and the theory of a chemosensory circuit in C. elegans　　　　　　　　　　　　　　　
M. Kuramochi; M. Doi
The Irago Conference 2014, Nov. 2014

Toward quantitative understanding of neuronal dynamics in C. elegans　　　　　　　　　　　　　　　
M. Kuramochi; M. Doi
System Neurobiology Spring School 2014 (SNSS2014), Mar. 2014

Mathematical analysis of a blue-green algae in Lake Kasumigaura　　　　　　　　　　　　　　　
Masahiro Kuramochi
OIST APYSEF, Sep. 2011, [Invited]

Dynamical model variables, calcium imaging and design of C. elegans　　　　　　　　　　　　　　　
M. Kuramochi; Y. Iwasaki
7th Asia Biophysics Association (ABA) Symposium & Annual Meeting of the Indian Biophysical Society (IBS), Jan. 2011, [Invited]

Quantitative Modeling of Neuronal Dynamics in C. elegans　　　　　　　　　　　　　　　
M. Kuramochi; Y. Iwasaki
ICONIP2010, Oct. 2010

Neuronal model including Ca2+ dynamics in C. elegans　　　　　　　　　　　　　　　
M. Kuramochi; Y. Iwasaki
4th East Asia C. elegans Meeting, Jul. 2010

Apr. 2024 - Present

2023 - Present

2021 - Present

2021 - Present

2021 - Present

2021 - Present

2021 - Present

2021 - Present

2023 - 2023

北米神経科学学会

THE JAPANESE SOCIETY FOR ARTIFICIAL INTELLIGENCE

THE MOLECULAR BIOLOGY SOCIETY OF JAPAN

JAPANESE NEURAL NETWORK SOCIETY

THE JAPAN NEUROSCIENCE SOCIETY

X線を用いた高速１分子運動解析による膜タンパク質オペランド計測の実現
基盤研究(B)
国立研究開発法人産業技術総合研究所
01 Apr. 2023 - 31 Mar. 2026

機械学習を用いたX線透過動画からの高分子材料の動態物性評価　　　　　　　　　　　　　　　
Apr. 2024 - Mar. 2025

先進情報解析を用いた物質動態のX線ブリンキング計測　　　　　　　　　　　　　　　
Apr. 2024 - Mar. 2025

凍結低温制御分子の構造特異機能の解明および個体丸ごと保存技術の開発　　　　　　　　　　　　　　　
Oct. 2022 - Mar. 2025

筑波大学拠点TRiSTAR「大学×国研×企業の連携によるトップランナー育成プログラム」　　　　　　　　　　　　　　　
Aug. 2022 - Mar. 2025

原子スケールの動的特性を検出する微分X線ブリンキング法の開発 ー生体分子から無機・有機・高分子材料までー　　　　　　　　　　　　　　　
Sep. 2022 - Mar. 2023

個体生物の細胞内反応場およびタンパク質分子の回転揺らぎピコメートルX線計測　　　　　　　　　　　　　　　
Apr. 2022 - Mar. 2023

生体内氷晶のＸ線ナノスケール観測に基づいた不凍タンパク質多細胞系凍結保存法の確立
Grant-in-Aid for Early-Career Scientists
The University of Tokyo
Apr. 2021 - Mar. 2023

時分割回折X線ブリンキングによるソフトクリスタル動的特性観測
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
The University of Tokyo
Apr. 2020 - Mar. 2022

線虫Ｃ．エレガンスにおける凝集タンパク質および分子夾雑場のX線１分子観察
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
The University of Tokyo
Apr. 2020 - Mar. 2022

線虫 C. エレガンスのシナプス伝達実測データに基づく理論モデル構築　　　　　　　　　　　　　　　
Feb. 2021 - Feb. 2022

Mechanism of antifreeze proteins for improving the freezing- and cold-tolerance in the nematode Caenorhabditis elegans
Grant-in-Aid for Early-Career Scientists
The University of Tokyo
Apr. 2019 - Mar. 2021

時分割回折X線ブリンキングによる線虫の不凍タンパク質動態の検出　　　　　　　　　　　　　　　
東京大学
Apr. 2020 - Feb. 2021

生きた線虫の１分子動態と細胞応答を同時取得する時分割Ｘ線回折／細胞イメージング法　　　　　　　　　　　　　　　
Apr. 2019 - Mar. 2020

蛍光生体イメージングと計算機シミュレーションによる線虫神経の時空間活動解析
Grant-in-Aid for JSPS Fellows
University of Tsukuba
Apr. 2016 - Mar. 2018

特開2022-019115, 特願2020-122717, Ｘ線を用いた物質評価方法及び物質評価装置
倉持昌弘, 佐々木裕次, 三尾和弘