研究活動情報

• 2022年06月 第22回日本蛋白質科学会, 若手奨励賞


• 2017年10月 The 38th Symposium on UltraSonic Electronics (USE2017), 奨励賞

  野井 健太郎


• 2015年09月 肥後医育振興会, 学術奨励賞

  野井 健太郎


• 2011年11月 9th International Conference on AAA Proteins, ポスター賞

  野井 健太郎


• 2011年11月 特定領域 タンパク質の社会, ポスター賞 優秀賞

  野井 健太郎

• Interfacial viscoelastic remodeling of epithelial monolayers analyzed using QCM-D

  Hirotaka Koyama, Tsubasa S. Matsui, Kentaro Noi, Naoki Kagawa, Shinji Deguchi

  Japan Society of Mechanical Engineers, 2026年, Journal of Biomechanical Science and Engineering

  [査読有り]

  研究論文（学術雑誌）


• Tppp3 determines basal body positioning and identity of respiratory cilia via microtubule assembly and sphingolipid homeostasis

  Takafumi Sakai, Masahiro Kawakita, Arashi Seki, Katsuyoshi Takaoka, Misaki Kurata, Junpei Torikai, Sohshiroh Atsumi, Kanji Yawata, Kentaro Noi, Yosuke Fukutani, Keiichi Noguchi, Hirofumi Kiyokawa, Mitsuru Morimoto, Eri Takeuchi, Katsura Minegishi, Yoshitsugu Aoki, Shoko Fujimura, Takayuki Nishizaka, Takanari Inoue, Kentaro Nagaoka, Hiroshi Tsugawa, Hisayoshi Hayashi, Hiroshi Ishiguro, Takuma Kozono, Ikuroh Ohsawa, Yasunori Fujita, Hiroaki Matsunami, Hiroshi Hamada, Kyosuke Shinohara

  Cilia are hair-like organelles that protrude from the cell surface. In mammals, tracheal multiciliated cells (MCCs) play an important role in elimination of hazardous microorganisms by driving a unidirectional mucus flow. Although uniform orientation of ciliary beating is critical for the unidirectional flow, it remains unknown how MCCs establish uniform orientations and maintain identities of hundreds of ciliary membranes. This study focuses on investigating the roles of Tubulin Polymerization Promoting Family Member 3 (Tppp3) in MCC function. We generated a Tppp3-deficient mouse ( Tppp3 ^{∆ex2-4/∆ex2-4} ; Tppp3 knockout (KO)) and found that the Tppp3 KO mouse exhibited cough and hyposmia phenotype. The loss of Tppp3 disrupted the apical microtubules (MTs) meshwork in the tracheal MCCs, leading to random orientation and alignment of basal bodies (BBs) of the motile cilia. Unexpectedly, aberrant ciliary membrane fusions occurred in the trachea of the Tppp3 KO mice. We examined the underlying molecular mechanism of the ciliary membrane fusion by isolating the tracheal cilium. Liquid Chromatography-Mass Spectrometry (LC–MS) analysis as well as pharmacological analysis revealed that hyperaccumulation of a long chain ceramide at the ciliary membrane caused the membrane fusion. In addition, sensory cilia formation was impaired in the olfactory sensory neuron of the Tppp3 KO mice. Due to the lack of Tppp3, dendritic MT assembly that underlies long-range migration of BBs toward the cell surface was impaired. These findings demonstrate that Tppp3, as well as the defined intracellular MT architecture, regulate proper orientation/subcellular positioning of BBs and the independency of individual motile cilium membranes.

  Proceedings of the National Academy of Sciences, 2025年12月, Proceedings of the National Academy of Sciences, 122(49) (49)

  [査読有り]

  研究論文（学術雑誌）


• Monitoring proliferation and material production of Cupriavidus necator H16 using cell count and volume measurement

  Kouhei Kamasaka, Naoki Abekawa, Koji Takeda, Kentaro Noi, Mami Matsuda, Keiji Matsumoto, Noboru Yumoto, Akihiko Kondo, Tomohisa Hasunuma

  Springer Science and Business Media LLC, 2025年08月, Scientific Reports, 15(1) (1)

  [査読有り]

  研究論文（学術雑誌）

  神戸大学リポジトリ（Kernel）へのリンク


• Molecular Characterization of the MoxR AAA+ ATPase of Synechococcus sp. Strain NKBG15041c

  Kota Mano, Kentaro Noi, Kumiko Oe, Takahiro Mochizuki, Ken Morishima, Rintaro Inoue, Masaaki Sugiyama, Keiichi Noguchi, Kyosuke Shinohara, Masafumi Yohda, Akiyo Yamada

  We isolated a stress-tolerance-related gene from a genome library of Synechococcus sp. NKBG15041c. The expression of the gene in E. coli confers resistance against various stresses. The gene encodes a MoxR AAA+ ATPase, which was designated SyMRP since it belongs to the MRP subfamily. The recombinant SyMRP showed weak ATPase activity and protected citrate synthase from thermal aggregation. Interestingly, the chaperone activity of SyMRP is ATP-dependent. SyMRP exists as a stable hexamer, and ATP-dependent conformation changes were not detected via analytical ultracentrifugation (AUC) or small-angle X-ray scattering (SAXS). Although the hexameric structure predicted by AlphaFold 3 was the canonical flat-ring structure, the structures observed by atomic force microscopy (AFM) and transmission electron microscopy (TEM) were not the canonical ring structure. In addition, the experimental SAXS profiles did not show a peak that should exist in the symmetric-ring structure. Therefore, SyMRP seems to form a hexameric structure different from the canonical hexameric structure of AAA+ ATPase.

  MDPI AG, 2024年09月, International Journal of Molecular Sciences, 25(18) (18), 9955 - 9955

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  研究論文（学術雑誌）


• Cellular Penetration and Intracellular Dynamics of Perfluorocarbon-Conjugated DNA/RNA as a Potential Means of Conditional Nucleic Acid Delivery.

  Masako Takatsu, Kunihiko Morihiro, Honoka Watanabe, Mizue Yuki, Takara Hattori, Kentaro Noi, Kohsuke Aikawa, Keiichi Noguchi, Masafumi Yohda, Takashi Okazoe, Akimitsu Okamoto

  Nucleic acid-based therapeutics represent a novel approach for controlling gene expression. However, a practical delivery system is required that overcomes the poor cellular permeability and intercellular instability of nucleic acids. Perfluorocarbons (PFCs) are highly stable structures that can readily traverse the lipid membrane of cells. Thus, PFC-DNA/RNA conjugates have properties that offer a potential means of delivering nucleic acid therapeutics, although the cellular dynamics of the conjugates remain unknown. Here, we performed systematic analysis of the cellular permeability of sequence-controlled PFC-DNA conjugates (N[PFC]n-DNA, n = 1,2,3,4,5) that can be synthesized by conventional phosphoramidite chemistry. We showed that DNA conjugates with two or more PFC-containing units (N[PFC]n≥2-DNA) penetrated HeLa cells without causing cellular damage. Imaging analysis along with quantitative flow cytometry analysis revealed that N[PFC]2-DNA rapidly passes through the cell membrane and is evenly distributed within the cytoplasm. Moreover, N[PFC]2-modified cyclin B1-targeting siRNA promoted gene knockdown efficacy of 30% compared with naked siRNA. A similar cell penetration without associated toxicity was consistent among the seven different human cell lines tested. These unique cellular environmental properties make N[PFC]2-DNA/RNA a potential nucleic acid delivery platform that can meet a wide range of applications.

  2023年12月, ACS chemical biology, 18(12) (12), 2590 - 2598, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Mechanistic characterization of disulfide bond reduction of an ERAD substrate mediated by cooperation between ERdj5 and BiP.

  Xiaohan Cai, Shogo Ito, Kentaro Noi, Michio Inoue, Ryo Ushioda, Yukinari Kato, Kazuhiro Nagata, Kenji Inaba

  Endoplasmic reticulum (ER)-associated degradation (ERAD) is a protein quality control process that eliminates misfolded proteins from the ER. DnaJ homolog subfamily C member 10 (ERdj5) is a protein disulfide isomerase (PDI) family member that accelerates ER-associated degradation (ERAD) by reducing disulfide bonds of aberrant proteins with the help of an ER-resident chaperone BiP. However, the detailed mechanisms by which ERdj5 acts in concert with BiP are poorly understood. In this study, we reconstituted an in vitro system that monitors ERdj5-mediated reduction of disulfide-linked J-chain oligomers, known to be physiological ERAD substrates. Biochemical analyses using purified proteins revealed that J-chain oligomers were reduced to monomers by ERdj5 in a stepwise manner via trimeric and dimeric intermediates, and BiP synergistically enhanced this action in an ATP-dependent manner. Single-molecule observations of ERdj5-catalyzed J-chain disaggregation using high-speed atomic force microscopy (HS-AFM) demonstrated the stochastic release of small J-chain oligomers through repeated actions of ERdj5 on peripheral and flexible regions of large J-chain aggregates. Using systematic mutational analyses, ERAD substrate disaggregation mediated by ERdj5 and BiP was dissected at the molecular level.

  2023年09月, The Journal of biological chemistry, 105274 - 105274, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Acceleration of amyloid fibril formation by multichannel sonochemical reactor

  Kentaro Noi, Kichitaro Nakajima, Keiichi Yamaguchi, Masatomo So, Kensuke Ikenaka, Hideki Mochizuki, Yuji Goto, Hirotsugu OGI

  Formation of amyloid fibrils of various amyloidogenic proteins is dramatically enhanced by ultrasound irradiation. For applying this phenomenon to the study of protein aggregation science and diagnosis of neurodegenerative diseases, a multichannel ultrasound irradiation system with individually adjustable ultrasound-irradiation conditions is necessary. Here, we develop a sonochemical reaction system, where an ultrasonic transducer is placed in each well of a 96-well microplate to perform ultrasonic irradiation of sample solutions under various conditions with high reproducibility, and applied it for studying amyloid-fibril formation of amyloid $\beta$, $\alpha$-synuclein, $\beta$2-microglobulin, and lysozyme. The results clearly show that our instrument is superior to conventional shaking method in terms of degree of acceleration and reproducibility of fibril formation reaction. The acceleration degree is controllable by controlling the driving voltage applied to each transducer. We have thus succeeded in developing a useful tool for the study of amyloid fibril formation in various proteins.

  IOP Publishing, 2021年12月, Japanese Journal of Applied Physics

  [査読有り]

  研究論文（学術雑誌）


• Disaggregation Behavior of Amyloid β Fibrils by Anthocyanins Studied by Total-Internal-Reflection-Fluorescence Microscopy Coupled with a Wireless Quartz-Crystal Microbalance Biosensor.

  Kentaro Noi, Kensuke Ikenaka, Hideki Mochizuki, Yuji Goto, Hirotsugu Ogi

  Amyloid fibrils are formed from various proteins, some of which cause the corresponding neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases. It has been reported that many compounds inhibit the formation of amyloid fibrils. Anthocyanins are flavonoid pigments present in fruits and vegetables, which are known to suppress symptoms related with Alzheimer's disease. However, the influence of anthocyanins on the amyloid fibril remains unclear. Here, we succeeded in the direct monitoring of the disaggregation reaction of single amyloid β (Aβ) fibrils by anthocyanins using total-internal-reflection-fluorescence microscopy with a quartz-crystal microbalance (TIRFM-QCM). It is found that the disassembly activity to the Aβ fibrils depends on the number of hydroxyl groups in six-membered ring B of anthocyanin, and only delphinidin-3-galactoside, possessing three hydroxyl groups there, shows high disassembly activity. Our results show the importance of the number of hydroxyl groups and demonstrate the usefulness of TIRFM-QCM as a powerful tool in studying interactions between amyloid fibrils and compounds.

  2021年08月, Analytical chemistry, 93(32) (32), 11176 - 11183, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Determining the domain-level reaction-diffusion properties of an actin-binding protein transgelin-2 within cells.

  Takumi Saito, Daiki Matsunaga, Tsubasa S Matsui, Kentaro Noi, Shinji Deguchi

  Proteins in cells undergo repeated binding to other molecules, thereby reducing the apparent extent of their intracellular diffusion. While much effort has been made to analytically decouple these combined effects of pure diffusion and chemical binding, it is difficult with conventional approaches to attribute the measured quantities to the nature of specific domains of the proteins. Motivated by the common goal in cell signaling research aimed at identifying the domains responsible for particular intermolecular interactions, here we describe a framework for determining the local physicochemical properties of cellular proteins associated with immobile scaffolds. To validate this new approach, we apply it to transgelin-2, an actin-binding protein whose intracellular dynamics remains elusive. We develop a fluorescence recovery after photobleaching (FRAP)-based framework, in which comprehensive combinations of domain-deletion mutants are created, and the difference among them in FRAP response is analyzed. We demonstrate that transgelin-2 in actin stress fibers (SFs) interacts with F-actin via two separate domains, and the chemical properties are determined for the individual domains. Its pure diffusion properties independent of the association to F-actin is also obtained. Our approach will thus be useful, as presented here for transgelin-2, in addressing the signaling mechanism of cellular proteins associated with SFs.

  2021年07月, Experimental cell research, 404(1) (1), 112619 - 112619, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Distinct roles and actions of protein disulfide isomerase family enzymes in catalysis of nascent-chain disulfide bond formation

  Chihiro Hirayama, Kodai Machida, Kentaro Noi, Tadayoshi Murakawa, Masaki Okumura, Teru Ogura, Hiroaki Imataka, Kenji Inaba

  The mammalian endoplasmic reticulum (ER) harbors more than 20 members of the protein disulfide isomerase (PDI) family that act to maintain proteostasis. Herein, we developed an in vitro system for directly monitoring PDI- or ERp46-catalyzed disulfide bond formation in ribosome-associated nascent chains of human serum albumin. The results indicated that ERp46 more efficiently introduced disulfide bonds into nascent chains with a short segment exposed outside the ribosome exit site than PDI. Single-molecule analysis by high-speed atomic force microscopy further revealed that PDI binds nascent chains persistently, forming a stable face-to-face homodimer, whereas ERp46 binds for a shorter time in monomeric form, indicating their different mechanisms for substrate recognition and disulfide bond introduction. Thus, ERp46 serves as a more potent disulfide introducer especially during the early stages of translation, whereas PDI can catalyze disulfide formation when longer nascent chains emerge out from ribosome.

  Elsevier BV, 2021年04月, iScience, 24(4) (4), 102296 - 102296, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Optimized sonoreactor for accelerative amyloid-fibril assays through enhancement of primary nucleation and fragmentation.

  Kichitaro Nakajima, Kentaro Noi, Keiichi Yamaguchi, Masatomo So, Kensuke Ikenaka, Hideki Mochizuki, Hirotsugu Ogi, Yuji Goto

  Ultrasonication to supersaturated protein solutions forcibly forms amyloid fibrils, thereby allowing the early-stage diagnosis for amyloidoses. Previously, we constructed a high-throughput sonoreactor to investigate features of the amyloid-fibril nucleation. Although the instrument substantiated the ultrasonication efficacy, several challenges remain; the key is the precise control of the acoustic field in the reactor, which directly affects the fibril-formation reaction. In the present study, we develop the optimized sonoreactor for the amyloid-fibril assay, which improves the reproducibility and controllability of the fibril formation. Using β2-microglobulin, we experimentally demonstrate that achieving identical acoustic conditions by controlling oscillation amplitude and frequency of each transducer results in identical fibril-formation behavior across 36 solutions. Moreover, we succeed in detecting the 100-fM seeds using the developed sonoreactor at an accelerated rate. Finally, we reveal that the acceleration of the fibril-formation reaction with the seeds is achieved by enhancing the primary nucleation and the fibril fragmentation through the analysis of the fibril-formation kinetics. These results demonstrate the efficacy of the developed sonoreactor for the diagnosis of amyloidoses owing to the accelerative seed detection and the possibility for further early-stage diagnosis even without seeds through the accelerated primary nucleation.

  2021年03月, Ultrasonics sonochemistry, 73, 105508 - 105508, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Split conformation of Chaetomium thermophilum Hsp104 disaggregase.

  Yosuke Inoue, Yuya Hanazono, Kentaro Noi, Akihiro Kawamoto, Masato Kimatsuka, Ryuhei Harada, Kazuki Takeda, Ryoichi Kita, Natsuki Iwamasa, Kyoka Shibata, Keiichi Noguchi, Yasuteru Shigeta, Keiichi Namba, Teru Ogura, Kunio Miki, Kyosuke Shinohara, Masafumi Yohda

  Hsp104 and its bacterial homolog ClpB form hexameric ring structures and mediate protein disaggregation. The disaggregated polypeptide is thought to thread through the central channel of the ring. However, the dynamic behavior of Hsp104 during disaggregation remains unclear. Here, we reported the stochastic conformational dynamics and a split conformation of Hsp104 disaggregase from Chaetomium thermophilum (CtHsp104) in the presence of ADP by X-ray crystallography, cryo-electron microscopy (EM), and high-speed atomic force microscopy (AFM). ADP-bound CtHsp104 assembles into a 65 left-handed spiral filament in the crystal structure at a resolution of 2.7 Å. The unit of the filament is a hexamer of the split spiral structure. In the cryo-EM images, staggered and split hexameric rings were observed. Further, high-speed AFM observations showed that a substrate addition enhanced the conformational change and increased the split structure's frequency. Our data suggest that split conformation is an off-pathway state of CtHsp104 during disaggregation.

  2021年02月, Structure (London, England : 1993), 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Visualization of structural dynamics of protein disulfide isomerase enzymes in catalysis of oxidative folding and reductive unfolding

  Masaki Okumura, Kentaro Noi, Kenji Inaba

  Time-resolved single-molecule observations by high-speed atomic force microscopy (HS-AFM), have greatly advanced our understanding of how proteins operate to fulfill their unique functions. Using this device, we succeeded in visualizing two members of the protein disulfide isomerase family (PDIs) that act to catalyze oxidative folding and reductive unfolding in the endoplasmic reticulum (ER). ERdj5, an ER-resident disulfide reductase that promotes ER-associated degradation, reduces nonnative disulfide bonds of misfolded proteins utilizing the dynamics of its N-terminal and C-terminal clusters. With unfolded substrates, canonical PDI assembles to form a face-to-face dimer with a central hydrophobic cavity and multiple redox-active sites to accelerate oxidative folding inside the cavity. Altogether, PDIs exert highly dynamic mechanisms to ensure the protein quality control in the ER.

  Elsevier BV, 2021年02月, Current Opinion in Structural Biology, 66, 49 - 57, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Mechanism of affinity-enhanced protein adsorption on bio-nanocapsules studied by viscoelasticity measurement with wireless QCM biosensor

  Kentaro Noi, Masumi Iijima, Shun'Ichi Kuroda, Fumihito Kato, Hirotsugu Ogi

  2020年07月, Japanese Journal of Applied Physics, 59(SK) (SK)

  [査読有り]

  研究論文（学術雑誌）


• TRANSIENT SPLITTING OF HSP104 HEXAMERIC RING AND ITS IMPLICATION IN PROTEIN DISAGGREGATION

  Yohda Masafumi, Inoue Yosuke, Hanazono Yuya, Noi Kentaro, Kawamoto Akihiro, Takeda Kazuki, Noguchi Keiichi, Namba Keiichi, Ogura Teru, Miki Kunio, Shinohara Kyosuke

  2019年09月, PROTEIN SCIENCE, 28, 22

  [査読有り]


• Ultrahigh-sensitive wireless QCM with bio-nanocapsules

  Kentaro Noi, Masumi Iijima, Shun'ichi Kuroda, Hirotsugu Ogi

  2019年08月, Sensors and Actuators, B: Chemical, 293, 59 - 62

  [査読有り]

  研究論文（学術雑誌）


• Ultrahigh-frequency, wireless mems qcm biosensor for direct, label-free detection of biomarkers in a large amount of contaminants

  Kentaro Noi, Arihiro Iwata, Fumihito Kato, Hirotsugu Ogi

  2019年08月, Analytical Chemistry, 91(15) (15), 9398 - 9402

  [査読有り]

  研究論文（学術雑誌）


• Cyclization of Single-Chain Fv Antibodies Markedly Suppressed Their Characteristic Aggregation Mediated by Inter-Chain VH-VL Interactions.

  Soichiro Yamauchi, Yoshihiro Kobashigawa, Natsuki Fukuda, Manaka Teramoto, Yuya Toyota, Chenjiang Liu, Yuka Ikeguchi, Takashi Sato, Yuko Sato, Hiroshi Kimura, Takeshi Masuda, Sumio Ohtsuki, Kentaro Noi, Teru Ogura, Hiroshi Morioka

  Single-chain Fv (scFv) antibodies are recombinant proteins in which the variable regions of the heavy chain (VH) and light chain (VL) are connected by a short flexible polypeptide linker. ScFvs have the advantages of easy genetic manipulation and low-cost production using Escherichia coli compared with monoclonal antibodies, and are thus expected to be utilized as next-generation medical antibodies. However, the practical use of scFvs has been limited due to low homogeneity caused by their aggregation propensity mediated by inter-chain VH-VL interactions. Because the interactions between the VH and VL domains of antibodies are generally weak, individual scFvs are assumed to be in equilibrium between a closed state and an open state, in which the VH and VL domains are assembled and disassembled, respectively. This dynamic feature of scFvs triggers the formation of dimer, trimer, and larger aggregates caused by the inter-chain VH-VL interactions. To overcome this problem, the N-terminus and C-terminus were herein connected by sortase A-mediated ligation to produce a cyclic scFv. Open-closed dynamics and aggregation were markedly suppressed in the cyclic scFv, as judged from dynamic light scattering and high-speed atomic force microscopy analyses. Surface plasmon resonance and differential scanning fluorometry analysis revealed that neither the affinity for antigen nor the thermal stability was disrupted by the scFv cyclization. Generality was confirmed by applying the present method to several scFv proteins. Based on these results, cyclic scFvs are expected to be widely utilized in industrial and therapeutic applications.

  2019年07月, Molecules (Basel, Switzerland), 24(14) (14), 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Dynamic assembly of protein disulfide isomerase in catalysis of oxidative folding

  Masaki Okumura, Kentaro Noi, Shingo Kanemura, Misaki Kinoshita, Tomohide Saio, Yuichi Inoue, Takaaki Hikima, Shuji Akiyama, Teru Ogura, Kenji Inaba

  Time-resolved direct observations of proteins in action provide essential mechanistic insights into biological processes. Here, we present mechanisms of action of protein disulfide isomerase (PDI)-the most versatile disulfide-introducing enzyme in the endoplasmic reticulum-during the catalysis of oxidative protein folding. Single-molecule analysis by high-speed atomic force microscopy revealed that oxidized PDI is in rapid equilibrium between open and closed conformations, whereas reduced PDI is maintained in the closed state. In the presence of unfolded substrates, oxidized PDI, but not reduced PDI, assembles to form a face-to-face dimer, creating a central hydrophobic cavity with multiple redox-active sites, where substrates are likely accommodated to undergo accelerated oxidative folding. Such PDI dimers are diverse in shape and have different lifetimes depending on substrates. To effectively guide proper oxidative protein folding, PDI regulates conformational dynamics and oligomeric states in accordance with its own redox state and the configurations or folding states of substrates.

  Springer Science and Business Media LLC, 2019年05月, Nature Chemical Biology, 15(5) (5), 499 - 509, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Ultrasonication-based rapid amplification of α-synuclein aggregates in cerebrospinal fluid.

  Keita Kakuda, Kensuke Ikenaka, Katsuya Araki, Masatomo So, César Aguirre, Yuta Kajiyama, Kuni Konaka, Kentaro Noi, Kousuke Baba, Hiroshi Tsuda, Seiichi Nagano, Takuma Ohmichi, Yoshitaka Nagai, Takahiko Tokuda, Omar M A El-Agnaf, Hirotsugu Ogi, Yuji Goto, Hideki Mochizuki

  α-Synuclein aggregates, a key hallmark of the pathogenesis of Parkinson's disease, can be amplified by using their seeding activity, and the evaluation of the seeding activity of cerebrospinal fluid (CSF) is reportedly useful for diagnosis. However, conventional shaking-based assays are time-consuming procedures, and the clinical significance of the diversity of seeding activity among patients remains to be clarified. Previously, we reported a high-throughput ultrasonication-induced amyloid fibrillation assay. Here, we adapted this assay to amplify and detect α-synuclein aggregates from CSF, and investigated the correlation between seeding activity and clinical indicators. We confirmed that this assay could detect α-synuclein aggregates prepared in vitro and also aggregates released from cultured cells. The seeding activity of CSF correlated with the levels of α-synuclein oligomers measured by an enzyme-linked immunosorbent assay. Moreover, the seeding activity of CSF from patients with Parkinson's disease was higher than that of control patients. Notably, the lag time of patients with Parkinson's disease was significantly correlated with the MIBG heart-to-mediastinum ratio. These findings showed that our ultrasonication-based assay can rapidly amplify misfolded α-synuclein and can evaluate the seeding activity of CSF.

  2019年04月, Scientific reports, 9(1) (1), 6001 - 6001, 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• A newly isolated Pex7-binding, atypical PTS2 protein P7BP2 is a novel dynein-type AAA+ protein

  Hajime Niwa, Yasuhiro Miyauchi-Nanri, Kanji Okumoto, Satoru Mukai, Kentaro Noi, Teru Ogura, Yukio Fujiki

  Oxford University Press (OUP), 2018年09月, The Journal of Biochemistry

  [査読有り]

  研究論文（学術雑誌）


• Expression, Functional Characterization, and Preliminary Crystallization of the Cochaperone Prefoldin from the Thermophilic Fungus Chaetomium thermophilum.

  Kento Morita, Yohei Y Yamamoto, Ayaka Hori, Tomohiro Obata, Yuko Uno, Kyosuke Shinohara, Keiichi Noguchi, Kentaro Noi, Teru Ogura, Kentaro Ishii, Koichi Kato, Mahito Kikumoto, Rocio Arranz, Jose M Valpuesta, Masafumi Yohda

  Prefoldin is a hexameric molecular chaperone found in the cytosol of archaea and eukaryotes. Its hexameric complex is built from two related classes of subunits, and has the appearance of a jellyfish: Its body consists of a double β-barrel assembly with six long tentacle-like coiled coils protruding from it. Using the tentacles, prefoldin captures an unfolded protein substrate and transfers it to a group II chaperonin. Based on structural information from archaeal prefoldins, mechanisms of substrate recognition and prefoldin-chaperonin cooperation have been investigated. In contrast, the structure and mechanisms of eukaryotic prefoldins remain unknown. In this study, we succeeded in obtaining recombinant prefoldin from a thermophilic fungus, Chaetomium thermophilum (CtPFD). The recombinant CtPFD could not protect citrate synthase from thermal aggregation. However, CtPFD formed a complex with actin from chicken muscle and tubulin from porcine brain, suggesting substrate specificity. We succeeded in observing the complex formation of CtPFD and the group II chaperonin of C. thermophilum (CtCCT) by atomic force microscopy and electron microscopy. These interaction kinetics were analyzed by surface plasmon resonance using Biacore. Finally, we have shown the transfer of actin from CtPFD to CtCCT. The study of the folding pathway formed by CtPFD and CtCCT should provide important information on mechanisms of the eukaryotic prefoldin⁻chaperonin system.

  2018年08月, International journal of molecular sciences, 19(8) (8), 英語, 国際誌

  [査読有り]

  研究論文（学術雑誌）


• Viscoelasticity Response during Fibrillation of Amyloid β Peptides on a Quartz-Crystal-Microbalance Biosensor

  Yen Ting Lai, Hirotsugu Ogi, Kentaro Noi, Fumihito Kato

  2018年05月, Langmuir, 34(19) (19), 5474 - 5479

  [査読有り]

  研究論文（学術雑誌）


• Production of Single-Chain Fv Antibodies Specific for GA-Pyridine, an Advanced Glycation End-Product (AGE), with Reduced Inter-Domain Motion

  Natsuki Fukuda, Kentaro Noi, Lidong Weng, Yoshihiro Kobashigawa, Hiromi Miyazaki, Yukari Wakeyama, Michiyo Takaki, Yusuke Nakahara, Yuka Tatsuno, Makiyo Uchida-Kamekura, Yoshiaki Suwa, Takashi Sato, Naoki Ichikawa-Tomikawa, Motoyoshi Nomizu, Yukio Fujiwara, Fumina Ohsaka, Takashi Saito, Katsumi Maenaka, Hiroyuki Kumeta, Shoko Shinya, Chojiro Kojima, Teru Ogura, Hiroshi Morioka

  2017年10月, MOLECULES, 22(10) (10), 英語

  [査読有り]

  研究論文（学術雑誌）


• RNA Aptamer Binds Heparin-Binding Epidermal Growth Factor-Like Growth Factor with High Affinity and Specificity and Neutralizes Its Activity

  Masaki Yamato, Takashi Matsuzaki, Ryo Araki, Shota Tsuchida, Keiji Okuda, Hai Ying Fu, Shoji Sanada, Hiroshi Asanuma, Yoshihiro Asano, Masanori Asakura, Hiroomi Torii, Kentaro Noi, Hirotsugu Ogi, Ryo Iwamoto, Eisuke Mekada, Seiji Takashima, Masafumi Kitakaze, Yasushi Sakata, Tetsuo Minamino

  2017年09月, INTERNATIONAL JOURNAL OF GERONTOLOGY, 11(3) (3), 191 - 196, 英語

  [査読有り]

  研究論文（学術雑誌）


• The Highly Dynamic Nature of ERdj5 Is Key to Efficient Elimination of Aberrant Protein Oligomers through ER-Associated Degradation

  Ken-ichi Maegawa, Satoshi Watanabe, Kentaro Noi, Masaki Okumura, Yuta Amagai, Michio Inoue, Ryo Ushioda, Kazuhiro Nagata, Teru Ogura, Kenji Inaba

  2017年06月, STRUCTURE, 25(6) (6), 846 - +, 英語

  [査読有り]

  研究論文（学術雑誌）


• A substrate for observing histidine-tagged proteins by high-speed atomic force microscopy

  Daisuke Yamamoto, Kentaro Noi, Teru Ogura

  福岡大学研究推進部, 2017年, Fukuoka University science reports, 47(1) (1), 1 - 6, 英語

  [査読有り]


• Nucleus factory on cavitation bubble for amyloid beta fibril

  Kichitaro Nakajima, Hirotsugu Ogi, Kanta Adachi, Kentaro Noi, Masahiko Hirao, Hisashi Yagi, Yuji Goto

  2016年02月, SCIENTIFIC REPORTS, 6, 英語

  [査読有り]

  研究論文（学術雑誌）


• Nucleation-fibrillation dynamics of A beta(1-40) peptides on liquid-solid surface studied by total-internal-reflection fluorescence microscopy coupled with quartz-crystal microbalance biosensor

  Hiroki Hamada, Hirotsugu Ogi, Kentaro Noi, Hisashi Yagi, Yuji Goto, Masahiko Hirao

  2015年07月, JAPANESE JOURNAL OF APPLIED PHYSICS, 54(7) (7), 英語

  [査読有り]

  研究論文（学術雑誌）


• Microtubule Severing by Katanin p60 AAA+ ATPase Requires the C-terminal Acidic Tails of Both α- and β-Tubulins and Basic Amino Acid Residues in the AAA+ Ring Pore

  Ai Johjima, Kentaro Noi, Shingo Nishikori, Hirotsugu Ogi, Masatoshi Esaki, Teru Ogura

  2015年05月, Journal of Biological Chemistry, 290(18) (18), 11762 - 11770

  [査読有り]

  研究論文（学術雑誌）


• Ultrafast propagation of beta-amyloid fibrils in oligomeric cloud

  Hirotsugu Ogi, Masahiko Fukukshima, Hiroki Hamada, Kentaro Noi, Masahiko Hirao, Hisashi Yagi, Yuji Goto

  2014年11月, SCIENTIFIC REPORTS, 4, 英語

  [査読有り]

  研究論文（学術雑誌）


• High-Speed Atomic Force Microscopic Observation of ATP-Dependent Rotation of the AAA plus Chaperone p97

  Kentaro Noi, Daisuke Yamamoto, Shingo Nishikori, Ken-ichi Arita-Morioka, Takayuki Kato, Toshio Ando, Teru Ogura

  2013年11月, STRUCTURE, 21(11) (11), 1992 - 2002, 英語

  [査読有り]

  研究論文（学術雑誌）


• Suppression of Sup35 amyloid fibril formation by group II chaperonin from Thermoplasma acidophilum

  Kentaro Noi, Aya Kitamura, Hidenori Hirai, Kunihiro Hngo, Toshihiko Sakurai, Tomohiro Mizobata, Yasushi Kawata

  Scientific Research Publishing, Inc., 2012年07月, American Journal of Molecular Biology, 2(3) (3), 265 - 275, 英語

  [査読有り]

  研究論文（学術雑誌）


• A Potentially Versatile Nucleotide Hydrolysis Activity of Group II Chaperonin Monomers from Thermoplasma acidophilum

  Kentaro Noi, Hidenori Hirai, Kunihiro Hongo, Tomohiro Mizobata, Yasushi Kawata

  2009年10月, BIOCHEMISTRY, 48(40) (40), 9405 - 9415, 英語

  [査読有り]

  研究論文（学術雑誌）


• Functional characterization of the recombinant group II chaperonin alpha from Thermoplasma acidophilum

  Hidenori Hirai, Kentaro Noi, Kunihiro Hongo, Tomohiro Mizobata, Yasushi Kawata

  2008年04月, JOURNAL OF BIOCHEMISTRY, 143(4) (4), 505 - 515, 英語

  [査読有り]

  研究論文（学術雑誌）

• アミロイド線維に対するHsp104の脱凝集活性の機能解析

  野井健太郎, 柴田京華, 小椋光, 養王田正文, 篠原恭介

  2023年, 日本蛋白質科学会年会プログラム・要旨集, 23rd (CD-ROM)


• Hsp104によるα-シヌクレインのアミロイド線維の脱凝集活性の機能解析

  野井健太郎, 柴田京華, 小椋光, 養王田正文, 篠原恭介

  2023年, 日本生化学会大会(Web), 96th


• 分子シャペロンHsp104によるアミロイド線維分解機構の動的解析

  野井健太郎, 柴田京香, 小椋光, 養王田正文, 篠原恭介

  2022年, 日本臨床ストレス応答学会大会抄録集, 16th


• 高速AFMを用いたHsp104によるアミロイド線維脱凝集の動的解析

  野井健太郎, 柴田京華, 小椋光, 養王田正文, 篠原恭介

  2022年, 日本生化学会大会(Web), 95th


• 超音波デバイス 無線QCM粘弾性計測によるBNC上の蛋白質補足時の親和性向上機構の解明

  野井健太郎, 荻博次

  2021年, 超音波Techno, 33(5) (5)


• TIRFM-QCM測定によるアミロイド線維の動的解析

  野井健太郎, 池中建介, 望月秀樹, 後藤祐児, 荻博次

  2021年, 日本生化学会大会(Web), 94th


• 高周波TIRFM-QCM測定によるアミロイド線維形成の解析

  野井健太郎

  2020年, 村田学術振興財団年報, (34) (34)


• TIRFM-QCMを用いたアントシアニンによるアミロイド線維分解の動的解析

  野井健太郎, 鎌田航平, 後藤祐児, 荻博次

  2019年, 日本細胞生物学会大会(Web), 71st


• TIRFM-QCMを用いたアミロイド線維形成および分解の動的解析

  野井健太郎, 池中建介, 望月秀樹, 後藤祐児, 荻博次

  2018年, 日本生化学会大会(Web), 91st


• マルチHANABIを用いたアミロイド線維形成促進の研究

  野井健太郎, 笹原健二, 宗正智, 池中建介, 望月秀樹, 後藤祐児, 荻博次

  2017年, 日本生化学会大会(Web), 90th


• TDP-43アミロイド線維に対する分子シャペロンVCP/p97の作用機構の解析

  野井健太郎, 野井健太郎, 小椋光, 小椋光

  2016年, 日本蛋白質科学会年会プログラム・要旨集, 16th


• 分子シャペロンVCP/p97のTDP-43及びαシヌクレインアミロイド線維への作用機構

  野井健太郎, 野井健太郎, 小椋光, 小椋光

  2016年, 日本生化学会大会(Web), 89th


• AAA型シャペロンVCP/p97の分子機構の高速原子間力顕微鏡による解明

  野井健太郎, 野井健太郎, 小椋光, 小椋光

  2015年, 日本蛋白質科学会年会プログラム・要旨集, 15th


• バイオナノカプセルを用いた無線QCMの高感度化

  野井健太郎, 飯嶋益巳, 黒田俊一, 荻博次, 平尾雅彦

  2014年, 日本蛋白質科学会年会プログラム・要旨集, 14th


• 分子シャペロンVCP/p97とTDP-43アミロイド線維の分子間相互作用の解析

  野井健太郎, 有田(森岡)健一, 小椋光

  2013年, 日本蛋白質科学会年会プログラム・要旨集, 13th


• 分子シャペロンVCP/p97とTDP-43アミロイド線維の相互作用の分子機構の解析

  野井健太郎, 有田(森岡)健一, 小椋光

  2012年, 日本生化学会大会(Web), 85th


• AAAファミリー蛋白質p97のATP依存的な構造変化の解析

  野井健太郎, 山本大輔, 錦織伸吾, 錦織伸吾, 有田健一, 安藤敏夫, 安藤敏夫, 小椋光

  2011年, 日本蛋白質科学会年会プログラム・要旨集, 11th


• 高速原子間力顕微鏡を用いたAAAシャペロニンp97のATP依存的構造変化の解析

  野井健太郎, 山本大輔, 錦織伸吾, 森岡健一, 安藤敏夫, 小椋光

  2010年, 生化学


• グループII型シャペロニンによるSup35NMのアミロイド線維形成抑制

  野井健太郎, 北村文, 平井秀憲, 本郷邦広, 溝端知宏, 河田康志

  2010年, 日本蛋白質科学会年会プログラム・要旨集, 10th


• 古細菌Thermoplasma acidophilum由来単量体シャペロニンの金属特異的新規機能の解析

  野井健太郎, 平井秀憲, 本郷邦広, 本郷邦広, 溝端知宏, 溝端知宏, 河田康志, 河田康志

  2009年, 生化学, 81(7) (7)


• グループII型シャペロニンによるアミロイド線維形成抑制効果

  野井健太郎, 北村文, 平井秀憲, 本郷邦宏, 本郷邦宏, 溝端知宏, 溝端知宏, 河田康志, 河田康志

  2009年, 日本蛋白質科学会年会プログラム・要旨集, 9th


• グループII型シャペロニンによるSup35NMのアミロイド線維形成抑制

  野井健太郎, 北村文, 平井秀憲, 本郷邦宏, 溝端知宏, 河田康志, 本郷邦宏, 溝端知宏, 河田康志

  2009年, 生化学


• 古細菌Thermoplasma acidophilum由来単量体シャペロニンの新規機能の解析

  野井健太郎, 平井秀憲, 本郷邦広, 本郷邦広, 溝端知宏, 溝端知宏, 河田康志, 河田康志

  2008年, 日本蛋白質科学会年会プログラム・要旨集, 8th


• 古細菌Thermoplasma acidophilum由来単量体シャペロニンのCo²⁺,Mn²⁺依存的新規機能

  野井健太郎, 平井秀憲, 本郷邦広, 溝端知宏, 河田康志

  2008年, 生化学, 80(7) (7)


• 古細菌Thermoplasma acidophilum由来単量体シャペロニンによるCo²⁺,Mn²⁺依存的な新規ヌクレオチド加水分解活性

  野井健太郎, 平井秀憲, 本郷邦宏, 溝端知宏, 河田康志

  2007年, 生化学


• Thermoplasma acidophilum由来単量体シャペロニンの新規ヌクレオチド加水分解活性

  野井健太郎, 野井健太郎, 平井秀憲, 平井秀憲, 本郷邦広, 本郷邦広, 溝端知宏, 溝端知宏, 河田康志, 河田康志

  2007年, 日本蛋白質科学会年会プログラム・要旨集, 7th

• 過酷環境に生きる好熱性真菌の脱凝集因子がヒトのアミロイド線維を壊す原理の解明

  野井 健太郎

  日本学術振興会, 科学研究費助成事業, 基盤研究(C), 東京農工大学, 2024年04月01日 - 2027年03月31日


• 変異型アクチンの試験管内バイオジェネシスとその分子機構の解析

  町田 幸大, 野井 健太郎

  日本学術振興会, 科学研究費助成事業, 基盤研究(B), 兵庫県立大学, 2021年04月01日 - 2025年03月31日

  本年度は、試験管内合成したヒトのベータアクチンタンパク質のN 末端にアルギニン化修飾を行うため、ヒトのアルギニン転移酵素（ATE1）をクローニングし、発現・精製を行った。このATE1 を添加したヒト因子由来再構成型タンパク質合成系で、野生型アクチン（N 末端DDD）をコードするPlasmid DNA を用いてアクチンタンパク質を合成した。比較対象として、N 末端にアルギニンを付加した変異型アクチン（N 末端RDDD）も合成した。これら合成したサンプルをウエスタンブロッティングにより解析した。その結果、野生型アクチン（N 末端DDD）の約40% が、N 末端アルギニン化アクチンと同じ移動度にシフトしていた。これはすなわち、ATE1 を添加した再構成型翻訳システム内で新規に合成された野生型アクチンの約40% がアルギニン化されたことを示す結果であった。また線維化したアクチンをLifeact-EGFPで蛍光標識し共焦点顕微鏡による解析を行った結果、ATE1存在下で合成したアクチンの方がATE1非存在下で合成したアクチンよりも線維が長いものが観察され、アクチンのN末端アルギニン化は線維化促進に寄与していることを示唆する結果が得らえた。以上の一連の結果は、ヒト因子由来再構成型タンパク質合成系でヒトのベータアクチンの合成、フォールディング、修飾をワンポットで再現できることを示すものであり、修飾酵素を活性のある状態で調製できさえすれば、アクチン以外のタンパク質においても応用できる可能性が高く、翻訳中あるいは翻訳後修飾を試験管内で解析するための画期的な研究ツールとなることが期待される。


• リング状AAA型シャペロンの基質への作用機構のin vitro系による解明

  小椋 光, 江崎 雅俊, 山中 邦俊, 野井 健太郎

  日本学術振興会, 科学研究費助成事業, 基盤研究(B), 熊本大学, 2016年04月01日 - 2019年03月31日

  AAA型シャペロンp97/Cdc48/CDC-48が20Sプロテアソームと会合して、Sod1など一群の基質タンパク質を分解することを明らかにした。Cdc48は、基質タンパク質をポアループ依存的にほどいて、20Sプロテアソームに送り込むという機能が推定された。酵母ミトコンドリアの形態と外膜タンパク質の代謝において、Cdc48の補因子Ubx2はミトコンドリア外膜の膜融合因子Fzo1の代謝を促進し、Ubp3はミトコンドリアの形態維持に必要であることを明らかにした。線虫UBXN-6は、後期エンドソーム形成に関わるCDC-48の補因子で、飢餓によって発現が誘導されることを明らかにした。