112 results on '"Fumiaki Hayashi"'
Search Results
2. 19F chemical library and 19F-NMR for a weakly bound complex structure
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Shoko Shinya, Ritsuko Katahira, Kyoko Furuita, Toshihiko Sugiki, Young-Ho Lee, Yoshikazu Hattori, Kohei Takeshita, Atsushi Nakagawa, Aoi Kokago, Ken-ichi Akagi, Muneki Oouchi, Fumiaki Hayashi, Takanori Kigawa, Midori Takimoto-Kamimura, Toshimichi Fujiwara, and Chojiro Kojima
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Pharmacology ,Organic Chemistry ,Drug Discovery ,Pharmaceutical Science ,Molecular Medicine ,Biochemistry - Abstract
19F-NMR analysis using the optimized 19F chemical library enables the modeling of the structure of the weakly bound protein–compound complex, overcoming the difficulty in fragment-based drug discovery.
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- 2022
3. Protein stabilization and refolding in a gigantic self-assembled cage
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Daishi Fujita, Yuya Fujii, Fumiaki Hayashi, Makoto Fujita, Ryoto Suzuki, Mayu Yamada, Akimasa Matsugami, Jing-Ke Weng, Takahiro Nakama, and Maho Yagi-Utsumi
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Chemistry ,General Chemical Engineering ,Biochemistry (medical) ,General Chemistry ,Biochemistry ,Protein tertiary structure ,Chaperonin ,Folding (chemistry) ,Materials Chemistry ,Biophysics ,Environmental Chemistry ,Molecule ,Nanobiotechnology ,Self-assembly ,Protein stabilization ,Macromolecule - Abstract
Summary Spatial isolation of molecules is often a powerful strategy for regulating their molecular behavior. Biological systems employ such mechanisms well; however, scientists have yet to rival nature, particularly for macromolecular substrates. We demonstrated that the encapsulation of a protein in a molecular cage with an open framework stabilizes the tertiary structure of the protein and improves its enzymatic activity. Particularly, when the three-dimensionally confined enzyme was exposed to an organic solvent, its half-life was prolonged 1,000-fold. Kinetic and spectroscopic analysis of the enzymatic reaction revealed that the key to this stability is the isolated space; this is reminiscent of chaperonins, which use their large internal cavities to assist the folding of client proteins. The single-molecule protein caging affords a new type of protein-based nanobiotechnology that accelerates molecular biology research as well as industrial applications.
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- 2021
4. Peripheral granular lymphocytopenia and dysmorphic leukocytosis as simple prognostic markers in COVID‐19
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Setsuko Marutani, Takashi Miida, Yosuke Iwasaki, Yuki Horiuchi, Akihiko Matsuzaki, Sachiko Matsuoka, Kaori Saito, Kimiko Kaniyu, Tomohiko Ai, Yoko Tabe, Kinya Uchihashi, Fumiaki Hayashi, and Kumiko Nishibe
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Erythrocyte Indices ,Male ,medicine.medical_specialty ,Leukocytosis ,Neutrophils ,Anemia ,Clinical Biochemistry ,Toxic granulation ,Cytoplasmic Granules ,Cell morphology ,Severity of Illness Index ,Gastroenterology ,Lymphopenia ,Internal medicine ,Image Processing, Computer-Assisted ,medicine ,Humans ,Lymphocyte Count ,Lymphocytes ,Cell Shape ,Aged ,Covid‐19 ,medicine.diagnostic_test ,SARS-CoV-2 ,business.industry ,Biochemistry (medical) ,morphological anomalies ,COVID-19 ,Complete blood count ,Red blood cell distribution width ,Original Articles ,Hematology ,General Medicine ,Middle Aged ,Prognosis ,peripheral blood ,medicine.disease ,Blood Cell Count ,blood cell counts ,Döhle bodies ,Female ,Original Article ,Neural Networks, Computer ,granular lymphocytes ,Lymphocytopenia ,medicine.symptom ,business - Abstract
Introduction Developing prognostic markers can be useful for clinical decision‐making. Peripheral blood (PB) examination is simple and basic that can be performed in any facility. We aimed to investigate whether PB examination can predict prognosis in coronavirus disease (COVID‐19). Methods Complete blood count (CBC) and PB cell morphology were examined in 38 healthy controls (HCs) and 40 patients with COVID‐19. Patients with COVID‐19, including 26 mild and 14 severe cases, were hospitalized in Juntendo University Hospital (Tokyo, Japan) between April 1 and August 6, 2020. PB examinations were performed using Sysmex XN‐3000 automated hematology analyzer and Sysmex DI‐60 employing the convolutional neural network‐based automatic image‐recognition system. Results Compared with mild cases, severe cases showed a significantly higher incidence of anemia, lymphopenia, and leukocytosis (P
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- 2021
5. Crystallographic and Physicochemical Analysis of Bovine and Human Teeth Using X-ray Diffraction and Solid-State Nuclear Magnetic Resonance
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Noriko Hiraishi, Tadamu Gondo, Yasushi Shimada, Robert Hill, and Fumiaki Hayashi
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Biomaterials ,solid-state nuclear magnetic resonance ,heteronuclear ,apatites ,Biomedical Engineering ,enamel ,dentin ,X-ray diffraction - Abstract
Dental research often uses bovine teeth as a substitute for human teeth. The aim of this study was to evaluate differences in the crystalline nanostructures of enamel and dentin between bovine and human teeth, using X-ray diffraction (XRD) and solid-state nuclear magnetic resonance (NMR). The crystallite size (crystallinity) and microstrains were analyzed using XRD with the Rietveld refinement technique and the Halder–Wagner method. The 31P and 1H NMR chemical environments were analyzed by two-dimensional (2D) 1H-31P heteronuclear-correlation (HETCOR) magic-angle spinning (MAS) NMR spectroscopy. Enamel had a greater crystallite size and fewer microstrains than dentin for both bovine and human teeth. When compared between the species, the bovine apatite had a smaller crystallite size with more microstrains than the human apatite for both dentin and enamel. The 2D HETCOR spectra demonstrated that a water-rich layer and inorganic HPO42− ions were abundant in dentin; meanwhile, the hydroxyl group in the lattice site was more dominant in enamel. A greater intensity of the hydroxyl group was detected in human than in bovine for both dentin and enamel. For 31P projections, bovine dentin and bovine enamel have wider linewidths than human dentin and human enamel, respectively. There are differences in the crystallite profile between human and bovine. The results of dental research should be interpreted with caution when bovine teeth are substituted for human teeth.
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- 2022
6. G-quadruplex-forming aptamer enhances the peroxidase activity of myoglobin against luminol
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Koji Sode, Fumiaki Hayashi, Kazunori Ikebukuro, Kanjana Khunathai, Hitoshi Kuno, Jinhee Lee, Nasa Savory, Akimasa Matsugami, Daiki Oshikawa, Kenta Nakama, Taiki Saito, Mana Kanazashi, Kaori Tsukakoshi, and Yasuko Yamagishi
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Luminescence ,AcademicSubjects/SCI00010 ,Aptamer ,Heme ,Biology ,G-quadruplex ,Substrate Specificity ,law.invention ,Luminol ,03 medical and health sciences ,chemistry.chemical_compound ,Chemical Biology and Nucleic Acid Chemistry ,law ,Genetics ,Computer Simulation ,Nuclear Magnetic Resonance, Biomolecular ,Peroxidase ,030304 developmental biology ,Chemiluminescence ,0303 health sciences ,Myoglobin ,SELEX Aptamer Technique ,030302 biochemistry & molecular biology ,Rational design ,Aptamers, Nucleotide ,G-Quadruplexes ,chemistry ,Biochemistry ,biology.protein - Abstract
Aptamers can control the biological functions of enzymes, thereby facilitating the development of novel biosensors. While aptamers that inhibit catalytic reactions of enzymes were found and used as signal transducers to sense target molecules in biosensors, no aptamers that amplify enzymatic activity have been identified. In this study, we report G-quadruplex (G4)-forming DNA aptamers that upregulate the peroxidase activity in myoglobin specifically for luminol. Using in vitro selection, one G4-forming aptamer that enhanced chemiluminescence from luminol by myoglobin's peroxidase activity was discovered. Through our strategy—in silico maturation, which is a genetic algorithm-aided sequence manipulation method, the enhancing activity of the aptamer was improved by introducing mutations to the aptamer sequences. The best aptamer conserved the parallel G4 property with over 300-times higher luminol chemiluminescence from peroxidase activity more than myoglobin alone at an optimal pH of 5.0. Furthermore, using hemin and hemin-binding aptamers, we demonstrated that the binding property of the G4 aptamers to heme in myoglobin might be necessary to exert the enhancing effect. Structure determination for one of the aptamers revealed a parallel-type G4 structure with propeller-like loops, which might be useful for a rational design of aptasensors utilizing the G4 aptamer-myoglobin pair.
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- 2021
7. Development of an evaluation model to determine disease severity in <scp>COVID</scp> ‐19 using basic laboratory markers
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Ryosuke Maki, Yuki Horiuchi, Fumiaki Hayashi, Shuko Nojiri, Ikki Takehara, Yosuke Iwasaki, Kazunori Miyake, Takashi Miida, Tomohiko Ai, and Yoko Tabe
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SARS-CoV-2 ,Biochemistry (medical) ,Clinical Biochemistry ,COVID-19 ,Humans ,Hematology ,General Medicine ,Severity of Illness Index ,Biomarkers - Published
- 2022
8. Nearly complete 1H, 13C and 15N chemical shift assignment of monomeric form of N-terminal domain of Nephila clavipes major ampullate spidroin 2
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Keiji Numata, Ali D. Malay, Fumiaki Hayashi, Akimasa Matsugami, and Nur Alia Oktaviani
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congenital, hereditary, and neonatal diseases and abnormalities ,0303 health sciences ,biology ,Chemistry ,Spidroin ,030303 biophysics ,Nephila clavipes ,Antiparallel (biochemistry) ,biology.organism_classification ,Biochemistry ,03 medical and health sciences ,Crystallography ,SILK ,Structural Biology ,Side chain ,Spider silk ,Protein secondary structure ,MASP1 ,030304 developmental biology - Abstract
Spider dragline silk is well recognized due to its excellent mechanical properties. Dragline silk protein mainly consists of two proteins, namely, major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2). The MaSp N-terminal domain (NTD) conformation displays a strong dependence on ion and pH gradients, which is crucial for the self-assembly behavior of spider silk. In the spider major ampullate gland, where the pH is neutral and concentration of NaCl is high, the NTD forms a monomer. In contrast, within the spinning duct, where pH becomes more acidic (to pH ~ 5) and the concentration of salt is low, NTD forms a dimer in antiparallel orientation. In this study, we report near-complete backbone and side chain chemical shift assignment of the monomeric form of NTD of MaSp2 from Nephila clavipes at pH 7 in the presence of 300 mM NaCl. Our NMR data demonstrate that secondary structure of monomeric form of NTD MaSp2 consists of five helix regions.
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- 2020
9. Retrospective analysis of serum albumin reduction after thoracoscopic esophagectomy for esophageal cancer
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Yumi Yamamoto, Chiaki Yamashita, Hitoshi Nakamura, Eriko Minami, Fumiaki Hayashi, Kengo Nishimura, Toshiaki Kurasako, and Masakazu Yamaoka
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medicine.medical_specialty ,biology ,business.industry ,medicine.medical_treatment ,Serum albumin ,Esophageal cancer ,medicine.disease ,Gastroenterology ,Internal medicine ,medicine ,Retrospective analysis ,biology.protein ,Thoracoscopic esophagectomy ,business ,Reduction (orthopedic surgery) - Published
- 2020
10. Molecular insights into the unusually promiscuous and catalytically versatile Fe(II)/α-ketoglutarate-dependent oxygenase SptF
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Huiping Zhang, Akihito Nonoyama, Ikuro Abe, Fumiaki Hayashi, Hui Tao, Takahiro Mori, Shuang Lyu, He-Ping Chen, and Shoukou Lee
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Oxygenase ,Stereochemistry ,Chemistry - Abstract
Non-heme iron and α-ketoglutarate-dependent (Fe/αKG) oxygenases catalyze various oxidative biotransformations. Due to their catalytic flexibility and high efficiency, Fe/αKG oxygenases have attracted keen attention for their application as biocatalysts. Here, we report the biochemical and structural characterizations of the unusually promiscuous and catalytically versatile Fe/αKG oxygenase SptF, involved in the biosynthesis of fungal meroterpenoid emervaridones. The in vitro analysis reveals that SptF catalyzes four continuous oxidation reactions, including hydroxylation, desaturation, epoxidation, and skeletal rearrangement. SptF exhibits extremely broad substrate specificity toward various meroterpenoids, and efficiently produced novel cyclopropane-ring-fused 5/3/5/5/6/6 and 5/3/6/6/6 scaffolds from terretonins. Moreover, SptF also hydroxylates steroids, including androsterone, testosterone, and progesterone, with different regiospecificities. Crystallographic and structure-based mutagenesis studies of SptF reveal the molecular basis of the enzyme reactions, and suggest that the malleability of the loop region contributes to the remarkable substrate promiscuity. SptF exhibits great potential as a promising biocatalyst for oxidation reactions.
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- 2021
11. Ion effects on the conformation and dynamics of repetitive domains of a spider silk protein: implications for solubility and β-sheet formation
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Fumiaki Hayashi, Akimasa Matsugami, Keiji Numata, and Nur Alia Oktaviani
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Kosmotropic ,Silk ,Beta sheet ,010402 general chemistry ,01 natural sciences ,Catalysis ,Ion ,Protein structure ,Chlorides ,Materials Chemistry ,Animals ,Spider silk ,Solubility ,Nuclear Magnetic Resonance, Biomolecular ,010405 organic chemistry ,Hydrogen bond ,Chemistry ,Sodium ,Metals and Alloys ,Hydrogen Bonding ,Spiders ,General Chemistry ,Hydrogen-Ion Concentration ,Recombinant Proteins ,0104 chemical sciences ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Chaotropic agent ,Ceramics and Composites ,Biophysics ,Protein Conformation, beta-Strand - Abstract
The effect of ions on the structure and dynamics of a spider silk protein is elucidated. Chaotropic ions prevent intra- and inter-molecular interactions on the repetitive domain, which are required to maintain the solubility, while kosmotropic ions promote hydrogen bond interactions in the glycine-rich region, which are a prerequisite for β-sheet formation.
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- 2019
12. Molecular mechanisms of cell cryopreservation with polyampholytes studied by solid-state NMR
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Fumiaki Hayashi, Robin Rajan, Suong-Hyu Hyon, Kazuaki Matsumura, and Toshio Nagashima
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chemistry.chemical_classification ,Osmotic shock ,Cryoprotectant ,02 engineering and technology ,Polymer ,Nuclear magnetic resonance spectroscopy ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,Cryopreservation ,0104 chemical sciences ,Membrane ,chemistry ,Solid-state nuclear magnetic resonance ,Mechanics of Materials ,TA401-492 ,Biophysics ,General Materials Science ,0210 nano-technology ,Glass transition ,Materials of engineering and construction. Mechanics of materials - Abstract
Polyampholytes are emerging macromolecular membrane non-penetrating cryoprotectants; however, the mechanism behind their cryopreservation remains unclear. Here, we investigated the mechanism using solid-state NMR spectroscopy. The polymer-chain dynamics and the water and ion mobilities in the presence of various membrane penetrating and non-penetrating cryoprotectants were monitored at low temperatures to mimic cryopreservation conditions. NMR experiments revealed that the water, Sodium-ion, and polymer-chain signals in a carboxylated poly-ʟ-lysine (COOH-PLL) solution broadened upon cooling, indicating increasingly restricted mobility and increased solution viscosity. Moreover, strong intermolecular interactions facilitated the COOH-PLL glass transition, trapping water and salt in the gaps of the reversible matrix, preventing intracellular ice formation and osmotic shock during freezing; this reduced cell stress is responsible for cryoprotection. This simple NMR technique enabled the correlation of the cryoprotective properties of polymers that operate through mechanisms different from those of current cryoprotectants, and will facilitate the future molecular design of cryoprotectants. Cellular cryopreservation is an important tool in modern biology. Here, previously reported polyampholyte cryoprotectants are studied by solid-state NMR, revealing the molecular mechanisms at play.
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- 2021
13. Brevisulcenals-A1 and A2, Sulfate Esters of Brevisulcenals, Isolated from the Red Tide Dinoflagellate
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Masayuki, Satake, Raku, Irie, Patrick T, Holland, D Tim, Harwood, Feng, Shi, Yoshiyuki, Itoh, Fumiaki, Hayashi, and Huiping, Zhang
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Magnetic Resonance Spectroscopy ,Molecular Structure ,Cell Survival ,Sulfates ,dinoflagellate ,marine polyether ,Article ,Mice ,Structure-Activity Relationship ,red tide incident ,Ethers, Cyclic ,Cell Line, Tumor ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Dinoflagellida ,Animals ,harmful algal bloom - Abstract
Two different types of polycyclic ether toxins, namely brevisulcenals (KBTs) and brevisulcatic acids (BSXs), produced by the red tide dinoflagellate Karenia brevisulcata, were the cause of a toxic incident that occurred in New Zealand in 1998. Four major components, KBT-F, -G, -H, and -I, shown to be cytotoxic and lethal in mice, were isolated from cultured K. brevisulcata cells, and their structures were elucidated by spectroscopic analyses. New analogues, brevisulcenal-A1 (KBT-A1) and brevisulcenal-A2 (KBT-A2), toxins of higher polarity than that of known KBTs, were isolated from neutral lipophilic extracts of bulk dinoflagellate culture extracts. The structures of KBT-A1 and KBT-A2 were elucidated as sulfated analogues of KBT-F and KBT-G, respectively, by NMR and matrix-assisted laser desorption/ionization tandem mass spectrometry (MALDI TOF/TOF), and by comparison with the spectra of KBT-F and KBT-G. The cytotoxicities of the sulfate analogues were lower than those of KBT-F and KBT-G.
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- 2020
14. Interactions of boron released from surface pre-reacted glass ionomer with enamel/dentin and its effect on pH
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Noriko Hiraishi, Mahmoud Sayed, Junji Tagami, Fumiaki Hayashi, and Robert G. Hill
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Multidisciplinary ,Enamel paint ,Chemistry ,Science ,Glass ionomer cement ,Preventive dentistry ,chemistry.chemical_element ,pH meter ,Article ,Demineralization ,stomatognathic diseases ,medicine.anatomical_structure ,Adsorption ,stomatognathic system ,visual_art ,Restorative dentistry ,visual_art.visual_art_medium ,Dentin ,medicine ,Medicine ,Fourier transform infrared spectroscopy ,Boron ,Nuclear chemistry - Abstract
This study investigated the interaction of borate ions released from surface pre-reacted glass ionomer (S-PRG) fillers with the biological apatites of enamel and dentin using solid-state (SS) magic-angle spinning nuclear magnetic resonance (MAS-NMR) spectroscopy analysis. We further evaluated the effect of borate ions on the pH change. Bovine enamel and dentin powder were submerged in S-PRG eluate (with borate ion concentration of 100 mM) for 3 h, then washed with deionized water (DW). The dried enamel and dentin specimens were used for 11B SS-NMR and Fourier transform infrared spectroscopy (FT-IR) analysis. Enamel and dentin blocks were treated with three solutions: DW (control), S-PRG eluent and borate solution (100 mM). The treated samples were submerged in the demineralization solution and the pH was measured using a pH meter daily for 6 days. The surfaces of the enamel and dentin blocks were then observed using field emission scanning electron microscopy (FE-SEM). SS-NMR analysis revealed the presence of adsorbed borate ions in the enamel and dentin samples in a tetra-coordinated form. The pH results demonstrated an increase in pH values in the S-PRG and borate groups. SEM images showed that the surfaces of the control group were demineralized, whereas the surfaces of the S-PRG and borate groups were intact. These results concluded that borate ions could be adsorbed to enamel and dentin in the tetra-coordinated form. Borate ions possess a buffer capacity which may help to protect the tooth structure against acid attacks and promote remineralization.
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- 2020
15. The origins of binding specificity of a lanthanide ion binding peptide
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Yoichi Hosokawa, Nobuhiro Ishida, Akimasa Matsugami, Fumiaki Hayashi, Nobuaki Kikkawa, and Takaaki Hatanaka
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0301 basic medicine ,Lanthanide ,Magnetic Resonance Spectroscopy ,Biophysics ,lcsh:Medicine ,Calorimetry ,Molecular Dynamics Simulation ,Crystallography, X-Ray ,010402 general chemistry ,Lanthanoid Series Elements ,01 natural sciences ,Article ,Dissociation (chemistry) ,Ion ,03 medical and health sciences ,Ion binding ,Cations ,Molecule ,lcsh:Science ,Binding selectivity ,Binding Sites ,Multidisciplinary ,Aqueous solution ,Ionic radius ,Molecular Structure ,Chemistry ,lcsh:R ,Water ,Computational biology and bioinformatics ,Coordination chemistry ,0104 chemical sciences ,Crystallography ,030104 developmental biology ,Thermodynamics ,lcsh:Q ,Structural biology ,Peptides ,Inorganic chemistry - Abstract
Lanthanide ions (Ln3+) show similar physicochemical properties in aqueous solutions, wherein they exist as + 3 cations and exhibit ionic radii differences of less than 0.26 Å. A flexible linear peptide lanthanide binding tag (LBT), which recognizes a series of 15 Ln3+, shows an interesting characteristic in binding specificity, i.e., binding affinity biphasically changes with an increase in the atomic number, and shows a greater than 60-fold affinity difference between the highest and lowest values. Herein, by combining experimental and computational investigations, we gain deep insight into the reaction mechanism underlying the specificity of LBT3, an LBT mutant, toward Ln3+. Our results clearly show that LBT3-Ln3+ binding can be divided into three, and the large affinity difference is based on the ability of Ln3+ in a complex to be directly coordinated with a water molecule. When the LBT3 recognizes a Ln3+ with a larger ionic radius (La3+ to Sm3+), a water molecule can interact with Ln3+ directly. This extra water molecule infiltrates the complex and induces dissociation of the Asn5 sidechain (one of the coordinates) from Ln3+, resulting in a destabilizing complex and low affinity. Conversely, with recognition of smaller Ln3+ (Sm3+ to Yb3+), the LBT3 completely surrounds the ions and constructs a stable high affinity complex. Moreover, when the LBT3 recognizes the smallest Ln3+, namely Lu3+, although it completely surrounds Lu3+, an entropically unfavorable phenomenon specifically occurs, resulting in lower affinity than that of Yb3+. Our findings will be useful for the design of molecules that enable the distinction of sub-angstrom size differences.
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- 2020
16. Inhibitory effects of sesquiterpene lactones from the Indonesian marine sponge Lamellodysidea cf. herbacea on bone morphogenetic protein-induced osteoblastic differentiation
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Ryuji Uchida, Henki Rotinsulu, Fumiaki Hayashi, Magie M. Kapojos, Wilmar Maarisit, Ohgi Takahashi, Deiske A. Sumilat, Defny S. Wewengkang, Hiroyuki Yamazaki, Delfly B. Abdjul, Satoshi Ohte, Hiroshi Tomoda, Huiping Zhang, Takenobu Katagiri, and Michio Namikoshi
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Stereochemistry ,Clinical Biochemistry ,Mutant ,Pharmaceutical Science ,Bone morphogenetic protein ,Sesquiterpene ,Sesquiterpene lactone ,01 natural sciences ,Biochemistry ,Cell Line ,chemistry.chemical_compound ,Lactones ,Mice ,Structure-Activity Relationship ,Drug Discovery ,Animals ,Molecular Biology ,chemistry.chemical_classification ,Osteoblasts ,biology ,Dose-Response Relationship, Drug ,Molecular Structure ,010405 organic chemistry ,Organic Chemistry ,Absolute configuration ,Cell Differentiation ,biology.organism_classification ,Alkaline Phosphatase ,0104 chemical sciences ,Porifera ,010404 medicinal & biomolecular chemistry ,Sponge ,chemistry ,Indonesia ,Bone Morphogenetic Proteins ,Molecular Medicine ,Alkaline phosphatase ,Two-dimensional nuclear magnetic resonance spectroscopy ,Sesquiterpenes - Abstract
A new unique sesquiterpene lactone, bicyclolamellolactone A (1), was isolated together with two known monocyclofarnesol-type sesquiterpenes, lamellolactones A (2) and B (3), from the Indonesian marine sponge Lamellodysidea sp. (cf. L. herbacea). The planar structure of 1 was assigned based on its spectroscopic data (1D and 2D NMR, HRESIMS, UV, and IR spectra). The relative and absolute configuration of 1 was determined by comparison of its calculated and experimental electronic circular dichroism spectra in combination with NOESY correlations. Compounds 1–3 inhibited bone morphogenic protein (BMP)-induced alkaline phosphatase activity in mutant BMP receptor-carrying C2C12 cells with IC50 values of 51, 4.6, and 20 μM, respectively.
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- 2020
17. Nearly complete
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Nur Alia, Oktaviani, Ali D, Malay, Akimasa, Matsugami, Fumiaki, Hayashi, and Keiji, Numata
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Nitrogen Isotopes ,Protein Domains ,Proton Magnetic Resonance Spectroscopy ,Animal Structures ,Animals ,Spiders ,Amino Acid Sequence ,Carbon-13 Magnetic Resonance Spectroscopy ,Hydrogen-Ion Concentration ,Fibroins ,Protein Structure, Secondary - Abstract
Spider dragline silk is well recognized due to its excellent mechanical properties. Dragline silk protein mainly consists of two proteins, namely, major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2). The MaSp N-terminal domain (NTD) conformation displays a strong dependence on ion and pH gradients, which is crucial for the self-assembly behavior of spider silk. In the spider major ampullate gland, where the pH is neutral and concentration of NaCl is high, the NTD forms a monomer. In contrast, within the spinning duct, where pH becomes more acidic (to pH ~ 5) and the concentration of salt is low, NTD forms a dimer in antiparallel orientation. In this study, we report near-complete backbone and side chain chemical shift assignment of the monomeric form of NTD of MaSp2 from Nephila clavipes at pH 7 in the presence of 300 mM NaCl. Our NMR data demonstrate that secondary structure of monomeric form of NTD MaSp2 consists of five helix regions.
- Published
- 2020
18. Distinct residual and disordered structures of alpha-synuclein analyzed by amide-proton exchange and NMR signal intensity
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Shiki Morita, Yuji Goto, Chiaki Nishimura, Akimasa Matsugami, Rina Okuwaki, Fumiaki Hayashi, and Iori Shinmura
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Magnetic Resonance Spectroscopy ,Mutant ,Biophysics ,Rigidity (psychology) ,010402 general chemistry ,Fibril ,Intrinsically disordered proteins ,01 natural sciences ,Biochemistry ,Oligomer ,Signal ,Protein Structure, Secondary ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Humans ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Temperature ,Parkinson Disease ,Amides ,Magnetic Resonance Imaging ,0104 chemical sciences ,Intrinsically Disordered Proteins ,Crystallography ,chemistry ,Structural change ,Mutation (genetic algorithm) ,Mutation ,alpha-Synuclein ,Mutant Proteins ,Protons - Abstract
The residual solution structures of two alpha-synuclein mutants, A30P and A53T, observed in family members of patients with Parkinson's disease were compared with that of wild-type by NMR. The A53T substitution had been shown to accelerate fibril formation of alpha-synuclein, whereas the A30P mutation has the negative and positive effects on the formation of the fibril and spherical oligomer, respectively. The remaining structure was analyzed via amide-proton exchange and signal intensity measurements using NMR. Amide-proton exchange was used for both the calculation of kex values and ratio of kex at different temperatures. Effects of the A30P (N-terminal region) mutation were observed at the C-terminal region as a more flexible structure, suggesting that long-range interactions exist between the N- and C-terminal regions in alpha-synuclein. In addition, the N-terminal region adopted a more rigid structure in the A53T and A30P mutants than in the wild-type. It was concluded that the structural change caused by the mutations is related to the formation of a beta-hairpin at the initiation site of the N-terminal core structure. Furthermore, the signal intensity was used to estimate the rigidity of the structure. Higher signal intensities were observed for A30P at the 112, 113, and 116 C-terminal residues, suggesting that this region adopts more flexible structure. The ratio of the intensities at different temperatures indicated more flexible or rigid structures in the N-terminal region of A30P than in that of wild-type. Thus, using different approaches and temperatures is a good method to analyze residual structure in intrinsically disordered proteins.
- Published
- 2019
19. Mirilactams C–E, Novel Polycyclic Macrolactams Isolated from Combined-Culture of Actinosynnema mirum NBRC 14064 and Mycolic Acid-Containing Bacterium
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Huiping Zhang, Shotaro Hoshino, Masahiro Ozeki, Takayoshi Awakawa, Ikuro Abe, Chin Piow Wong, Hiroyuki Morita, Hiroyasu Onaka, and Fumiaki Hayashi
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chemistry.chemical_classification ,Circular dichroism ,Tsukamurella pulmonis ,Natural product ,biology ,010405 organic chemistry ,Stereochemistry ,General Chemistry ,General Medicine ,010402 general chemistry ,Polyene ,biology.organism_classification ,medicine.disease_cause ,01 natural sciences ,0104 chemical sciences ,Mycolic acid ,Actinobacteria ,chemistry.chemical_compound ,chemistry ,Biosynthesis ,Drug Discovery ,medicine ,Bacteria - Abstract
Mycolic acid-containing bacteria (MACB) are known to activate cryptic natural product biosynthesis in co-cultures with actinobacteria. We cultured Actinosynnema mirum NBRC 14064, a producer of the mono-cyclic polyene macrolactam mirilactam A (6), with the MACB Tsukamurella pulmonis TP-B0596. As a result, three novel compounds (mirilactams C-E, 1-3) were produced in the co-culture conditions. Compounds 1-3 were likely derived from 6 by epoxidation and subsequent spontaneous cyclization. The chemical structures and stereochemistries of 1-3 were determined by spectroscopic analyses (NMR and MS), conformational searches in the optimized potentials for liquid simulations-3 (OPLS3) force field, and calculations of electronic circular dichroism (ECD).
- Published
- 2018
20. Two new pyrrolo-2-aminoimidazoles from a Myanmarese marine sponge, Clathria prolifera
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Ikuro Abe, Hiroyuki Morita, So-Yeun Woo, Huiping Zhang, Aung Aung Aye, Nwet Nwet Win, Takuya Ito, Hla Ngwe, Shotaro Hoshino, Fumiaki Hayashi, Chin Piow Wong, and Nang Mya Han
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Antifungal ,Molecular Structure ,biology ,010405 organic chemistry ,Astrosclera willeyana ,Chemistry ,medicine.drug_class ,Stereochemistry ,Imidazoles ,Diastereomer ,Axinella ,010402 general chemistry ,biology.organism_classification ,01 natural sciences ,Porifera ,0104 chemical sciences ,Sponge ,medicine ,Animals ,Molecular Medicine ,Manzacidin D ,Enantiomer ,Clathria prolifera - Abstract
Marine organisms such as marine sponges and soft corals are valuable sources of pharmacologically active secondary metabolites. In our ongoing research on the discovery of new secondary metabolites from marine organisms, two new pyrrolo-2-aminoimidazoles, clathriroles A (1) and B (2), were isolated from the water-soluble portion prepared from the methanol and acetone (2:1) extract of the marine sponge, Clathria prolifera, collected in Myanmar. The chemical structures of the isolated compounds were determined using extensive spectroscopic techniques, including NMR, HRESIMS, IR, and optical rotation, and comparisons with the reported literature. The spectroscopic analyses of 1 and 2 suggested that 1 is an enantiomer of antifungal N-methylmanzacidin C isolated from the marine sponge Axinella brevistyla, whereas 2 is a diastereomer of manzacidin D at C-11 isolated from the marine sponge Astrosclera willeyana. To the best of our knowledge, this is the first report of the isolation of the pyrrolo-2-aminoimidazole compounds from C. prolifera. Furthermore, in contrast to the potency of N-methylmanzacidin C against Saccharomyces cerevisiae, the antifungal assay revealed that 1 and 2 lack any activity against this strain. Thus, these observations may suggest that the absolute configurations at both C-9 and C-11 play an important role in controlling the antifungal activity of this type of compound.
- Published
- 2018
21. Umezawamides, new bioactive polycyclic tetramate macrolactams isolated from a combined-culture of Umezawaea sp. and mycolic acid-containing bacterium
- Author
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Shotaro Hoshino, Fumiaki Hayashi, Shumpei Asamizu, Ikuro Abe, Takayoshi Awakawa, Huiping Zhang, Chin Piow Wong, Hiroyasu Onaka, and Masahiro Ozeki
- Subjects
0301 basic medicine ,Antifungal Agents ,Lactams, Macrocyclic ,Molecular Conformation ,Growth inhibitory ,Microbial Sensitivity Tests ,medicine.disease_cause ,Mycolic acid ,Mice ,03 medical and health sciences ,Cell Line, Tumor ,Actinomycetales ,Candida albicans ,Drug Discovery ,Bacteriology ,medicine ,Animals ,Humans ,Bioorganic chemistry ,Polycyclic Compounds ,Pharmacology ,chemistry.chemical_classification ,Tsukamurella pulmonis ,Antibiotics, Antineoplastic ,Bacteria ,biology ,Leukemia P388 ,Chemistry ,biology.organism_classification ,Anti-Bacterial Agents ,Genus Umezawaea ,030104 developmental biology ,Mycolic Acids ,Biochemistry - Abstract
New polycyclic tetramate macrolactams, Umezawamides A (1) and B (2) were isolated from a combined-culture of Umezawaea sp. RD066910 and mycolic-acid containing bacterium Tsukamurella pulmonis TP-B0596. Their planar structures and partial stereochemistries were determined based on the spectroscopic analysis, MMFF conformational search, and ECD calculations. Umezawamides are the first secondary metabolites isolated from the genus Umezawaea and they exhibited cytotoxicities to P388 murine leukemia cells. Furthermore, umezawamide A (1) showed growth inhibitory activity against Candida albicans.
- Published
- 2018
22. Beijinchromes A–D, Novel Aromatic Compounds Isolated from Nocardia beijingensis NBRC 16342
- Author
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Ikuro Abe, Fumiaki Hayashi, Huiping Zhang, Shotaro Hoshino, and Takayoshi Awakawa
- Subjects
Chromatography ,Antioxidant ,biology ,medicine.drug_class ,Chemistry ,Silica gel ,medicine.medical_treatment ,Antibiotics ,Nocardia ,General Chemistry ,General Medicine ,Reversed-phase chromatography ,Secondary metabolite ,biology.organism_classification ,chemistry.chemical_compound ,Drug Discovery ,medicine ,Nocardia beijingensis ,Gene ,medicine.drug - Abstract
Nocardia is a potent bacterial producer of bioactive compounds. From a culture of Nocardia beijingensis NBRC 16342, we isolated four aromatic compounds, named beijinchromes A-D (1-4). We purified them by silica gel chromatography and reverse phase HPLC, and identified their structures by NMR and high resolution (HR)-MS analyses. 1, 2, and 4 are novel 1,2,3,8-tetrasubstituted naphthalenes, and 3 is a novel 3,8-disubstituted ortho-naphthoquinone. 1 and 2 exert antioxidant activities, and 3 exhibits antibiotic activity. Remarkably, the putative biosynthetic gene clusters for 1-4 are widely distributed in 37 Nocardia species, implying their potential to produce this family of compounds and important biological functions of beijinchromes.
- Published
- 2019
23. Structures of the Largest Amphidinol Homologues from the Dinoflagellate Amphidinium carterae and Structure–Activity Relationships
- Author
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Huiping Zhang, Jong Souk Kim, Raymond S. Malabed, Fumiaki Hayashi, Shinya Hanashima, Jong-Soo Lee, Kimberly Cornelio, Michio Murata, Shoko Mori, Chang Hoon Kim, Masayuki Satake, and Nobuaki Matsumori
- Subjects
Antifungal ,Antifungal Agents ,medicine.drug_class ,ved/biology.organism_classification_rank.species ,Pharmaceutical Science ,Alkenes ,Biology ,010402 general chemistry ,Hemolysis ,01 natural sciences ,Analytical Chemistry ,Structure-Activity Relationship ,Polyketide ,Japan ,Polyol ,Amphidinium carterae ,Drug Discovery ,medicine ,Animals ,Structure–activity relationship ,Nuclear Magnetic Resonance, Biomolecular ,Pyrans ,Pharmacology ,chemistry.chemical_classification ,Molecular Structure ,010405 organic chemistry ,ved/biology ,Organic Chemistry ,Dinoflagellate ,biology.organism_classification ,0104 chemical sciences ,Complementary and alternative medicine ,chemistry ,Biochemistry ,Polyketides ,Dinoflagellida ,Molecular Medicine ,Drugs, Chinese Herbal - Abstract
Amphidinols are polyketide metabolites produced by marine dinoflagellates and are chiefly composed of a long linear chain with polyol groups and polyolefins. Two new homologues, amphidinols 20 (AM20, 1) and 21 (AM21, 2), were isolated from Amphidinium carterae collected in Korea. Their structures were elucidated by detailed NMR analyses as amphidinol 6-type compounds with remarkably long polyol chains. Amphidinol 21 (2) has the longest linear structure among the amphidinol homologues reported so far. The congeners, particularly amphidinol 21 (2), showed weaker activity in hemolysis and antifungal assays compared to known amphidinols.
- Published
- 2017
24. Designing the optimal public pension system
- Author
-
Jun Iritani, Fumiaki Hayashi, Kazumasa Oguro, and Takao Fujii
- Subjects
Consumption (economics) ,Actuarial science ,Order (exchange) ,Homogeneous ,Partial equilibrium ,Public pension ,Economics ,Econometrics ,Population growth ,Overlapping generations model ,General Economics, Econometrics and Finance - Abstract
This paper uses a two-period overlapping generations model in order to provide a theoretical design for the optimal public pension system based on a partial equilibrium analysis. Household preferences depend on two periods' consumption and leisure and are positively homogeneous of degree m with respect to consumption in the working and retired periods. We present characteristic features of the optimal public pension system in this paper. First, differences in the population growth rate do not affect the large/small relation between the optimal net lifetime burden rates of generations. Second, we present the optimal public pension system explicitly if m
- Published
- 2017
25. Beijinchromes A-D, Novel Aromatic Compounds Isolated from Nocardia beijingensis NBRC 16342
- Author
-
Shotaro, Hoshino, Takayoshi, Awakawa, Huiping, Zhang, Fumiaki, Hayashi, and Ikuro, Abe
- Subjects
Molecular Structure ,Stereoisomerism ,Naphthalenes ,Nocardia ,Naphthoquinones - Abstract
Nocardia is a potent bacterial producer of bioactive compounds. From a culture of Nocardia beijingensis NBRC 16342, we isolated four aromatic compounds, named beijinchromes A-D (1-4). We purified them by silica gel chromatography and reverse phase HPLC, and identified their structures by NMR and high resolution (HR)-MS analyses. 1, 2, and 4 are novel 1,2,3,8-tetrasubstituted naphthalenes, and 3 is a novel 3,8-disubstituted ortho-naphthoquinone. 1 and 2 exert antioxidant activities, and 3 exhibits antibiotic activity. Remarkably, the putative biosynthetic gene clusters for 1-4 are widely distributed in 37 Nocardia species, implying their potential to produce this family of compounds and important biological functions of beijinchromes.
- Published
- 2019
26. Lignans with melanogenesis effects from Premna serratifolia wood
- Author
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Huiping Zhang, Hiroyuki Morita, So-Yeun Woo, Ikuro Abe, Prema, Chin Piow Wong, Nwet Nwet Win, Maurice D. Awouafack, Hla Ngwe, Shotaro Hoshino, and Fumiaki Hayashi
- Subjects
Circular dichroism ,Stereochemistry ,Phytochemicals ,Melanoma, Experimental ,Infrared spectroscopy ,Myanmar ,01 natural sciences ,Lignans ,Melanin ,chemistry.chemical_compound ,Mice ,Cell Line, Tumor ,Drug Discovery ,Premna serratifolia ,Animals ,Cytotoxicity ,Pharmacology ,Lignan ,Melanins ,Lamiaceae ,biology ,Molecular Structure ,010405 organic chemistry ,General Medicine ,Mouse Melanoma ,biology.organism_classification ,Antineoplastic Agents, Phytogenic ,Wood ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,chemistry ,Two-dimensional nuclear magnetic resonance spectroscopy - Abstract
Three new lignoids, premnan A (1), premnan B (2), and tauntangyiol C (3), were isolated from Premna serratifolia wood, a traditional cosmetic plant in Myanmar, together with a new lignoid, premnan C (4) assumed to be an artifact, one natural new lignoid (5), and three known lignoids (6–8). The structures of the new compounds 1–4 were elucidated based on 1D and 2D NMR, IR spectroscopy, and HRESIMS. The absolute configurations of 1–4 were also determined by optical rotation, circular dichroism (CD) data analyses, and comparisons with the reported literature. All isolated compounds were tested for their melanogenesis activities against the B16-F10 mouse melanoma cell line. Compounds 1 and 4 showed melanogenesis enhancing activities of 31% and 50%, respectively, at a 50 μM concentration. Compounds 2, 3, and 6 increased melanin production by 67%, 30%, and 45%, respectively, at a 100 μM concentration, without any cytotoxicity.
- Published
- 2018
27. Genome-Based Discovery of an Unprecedented Cyclization Mode in Fungal Sesterterpenoid Biosynthesis
- Author
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Fumiaki Hayashi, Bin Qin, Takahiro Mori, Kazuya Nakagawa, Zhiyang Quan, Huiping Zhang, Masahiro Okada, Ikuro Abe, Takaaki Mitsuhashi, Hiroshi Kawaide, Yudai Matsuda, and Shotaro Hoshino
- Subjects
Sesterterpenes ,Stereochemistry ,Aspergillus oryzae ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Genome ,Catalysis ,Terpene ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Biosynthesis ,Geranylfarnesyl pyrophosphate ,Phylogeny ,Carbon chain ,Biological Products ,Alkyl and Aryl Transferases ,Emericella ,ATP synthase ,biology ,Terpenes ,010405 organic chemistry ,General Chemistry ,Carbon ,Recombinant Proteins ,Terpenoid ,0104 chemical sciences ,Diphosphates ,chemistry ,Cyclization ,biology.protein ,Genome, Fungal ,Hydrogen - Abstract
Sesterterpenoids are a group of terpenoid natural products that are primarily biosynthesized via cyclization of the C25 linear substrate geranylfarnesyl pyrophosphate (GFPP). Although the long carbon chain of GFPP in theory allows for many different cyclization patterns, sesterterpenoids are relatively rare species among terpenoids, suggesting that many intriguing sesterterpenoid scaffolds have been overlooked. Meanwhile, the recent identification of the first sesterterpene synthase has allowed the discovery of new sesterterpenoids by the genome mining approach. In this study, we characterized the unusual fungal sesterterpene synthase EvQS and successfully obtained the sesterterpene quiannulatene (1) with a novel and unique highly congested carbon skeleton, which is further oxidized to quiannulatic acid (2) by the cytochrome P450 Qnn-P450. A mechanistic study of its cyclization from GFPP indicated that the biosynthesis employs an unprecedented cyclization mode, which involves three rounds of hydride shifts and two successive C-C bond migrations to construct the 5-6-5-5-5 fused ring system of 1.
- Published
- 2016
28. Structural Analysis of the End Groups and Substructures of Commercial Poly(ethylene terephthalate) by Multiple-WET 1H/13C NMR
- Author
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Hiroshi Waki, Kimiko Tanaka, Muneki Oouchi, Hideaki Maeda, and Fumiaki Hayashi
- Subjects
Materials science ,Polymers and Plastics ,Organic Chemistry ,02 engineering and technology ,Carbon-13 NMR ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Inorganic Chemistry ,Polymer chemistry ,Materials Chemistry ,0210 nano-technology ,Poly ethylene - Published
- 2016
29. Astellifadiene: Structure Determination by NMR Spectroscopy and Crystalline Sponge Method, and Elucidation of its Biosynthesis
- Author
-
Yudai Matsuda, Fumiaki Hayashi, Huiping Zhang, Takahiro Mori, Ikuro Abe, Makoto Fujita, Takaaki Mitsuhashi, Manabu Hoshino, Masahiro Okada, and Shoukou Lee
- Subjects
Magnetic Resonance Spectroscopy ,Sesterterpenes ,Stereochemistry ,Aspergillus oryzae ,Molecular Conformation ,Stereoisomerism ,Sequence (biology) ,Crystallography, X-Ray ,010402 general chemistry ,01 natural sciences ,Catalysis ,chemistry.chemical_compound ,Emericella ,Biosynthesis ,chemistry.chemical_classification ,Biological Products ,Alkyl and Aryl Transferases ,biology ,010405 organic chemistry ,General Medicine ,General Chemistry ,Nuclear magnetic resonance spectroscopy ,biology.organism_classification ,Terpenoid ,0104 chemical sciences ,Hydrocarbon ,chemistry - Abstract
Genome mining of a terpene synthase gene from Emericella variecolor NBRC 32302 and its functional expression in Aspergillus oryzae led to the production of the new sesterterpene hydrocarbon, astellifadiene (1), having a 6-8-6-5-fused ring system. The structure of 1 was initially investigated by extensive NMR analyses, and was further confirmed by the crystalline sponge method, which established the absolute structure of 1 and demonstrated the usefulness of the method in the structure determination of complex hydrocarbon natural products. Furthermore, the biosynthesis of 1 was proposed on the basis of isotope-incorporation experiments performed both in vivo and in vitro. The cyclization of GFPP involves a protonation-initiated second cyclization sequence, 1,2-alkyl migration, and 1,5-hydride shift to generate the novel scaffold of 1.
- Published
- 2016
30. Dextran oxidized by a malaprade reaction shows main chain scission through a maillard reaction triggered by schiff base formation between aldehydes and amines
- Author
-
Suong-Hyu Hyon, Fumiaki Hayashi, Naoki Nakajima, Wichchulada Chimpibul, Kazuaki Matsumura, and Toshio Nagashima
- Subjects
Reaction mechanism ,Schiff base ,Polymers and Plastics ,Organic Chemistry ,Periodate ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Gel permeation chromatography ,chemistry.chemical_compound ,Maillard reaction ,symbols.namesake ,Dextran ,chemistry ,Polymer chemistry ,Materials Chemistry ,symbols ,Organic chemistry ,Hemiacetal ,Amine gas treating ,0210 nano-technology - Abstract
Although periodate-oxidized dextran is widely used in biomedical applications, the degradation mechanism of oxidized dextran has not yet been elucidated. Herein, we propose a novel main chain scission mechanism of oxidized dextran triggered by reaction with amine. NMR analysis revealed four hemiacetal substructures during oxidation by periodate. Kinetic analysis showed that the degradation time constant of the C3-removed substructure and increasing time constant of the reducing end protons are consistent with the decrease in molecular weight determined by gel permeation chromatography. A methylene group is generated at the same time constant of degradation, indicating that oxidized dextran degradation proceeds via a Maillard reaction. Oxidized dextran does not degrade in saline solution without reactive amine species. Thus, we conclude that oxidized dextran is degraded in the main chain via Schiff base formation through a Maillard reaction, depending on the oxidation ratio and amino acid concentration. These findings help to elucidate the reaction mechanism of polysaccharide degradation and develop novel biodegradable polysaccharide materials for biomedical applications. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016, 54, 2254–2260
- Published
- 2016
31. Niizalactams A–C, Multicyclic Macrolactams Isolated from Combined Culture of Streptomyces with Mycolic Acid-Containing Bacterium
- Author
-
Masahiro Okada, Ikuro Abe, Shotaro Hoshino, Huiping Zhang, Fumiaki Hayashi, Hiroyasu Onaka, and Toshiyuki Wakimoto
- Subjects
Stereochemistry ,Lactams, Macrocyclic ,Saccharomyces cerevisiae ,Pharmaceutical Science ,Microbial Sensitivity Tests ,Bacillus subtilis ,Biology ,medicine.disease_cause ,Streptomyces ,Analytical Chemistry ,Mycolic acid ,chemistry.chemical_compound ,Candida albicans ,Drug Discovery ,medicine ,Animals ,Derivatization ,Nuclear Magnetic Resonance, Biomolecular ,Pharmacology ,chemistry.chemical_classification ,Tsukamurella pulmonis ,Molecular Structure ,Leukemia P388 ,Organic Chemistry ,biology.organism_classification ,Mycolic Acids ,Complementary and alternative medicine ,chemistry ,Intramolecular force ,Molecular Medicine ,Drug Screening Assays, Antitumor ,Bacteria - Abstract
A terrestrial bacterium, Streptomyces sp. NZ-6, produced niizalactams A-C (1-3), unprecedented di- and tricyclic macrolactams, by coculturing with the mycolic acid-containing bacterium Tsukamurella pulmonis TP-B0596. Their complete structures, including absolute configurations, were elucidated on the basis of spectroscopic data and chemical derivatization. Their unique skeletons are proposed to be biosynthesized from a common 26-membered macrolactam intermediate by SN2 cyclization or an intramolecular Diels-Alder reaction.
- Published
- 2015
32. Inhibition of cholesteryl ester synthesis by polyacetylenes from Atractylodes rhizome
- Author
-
Huiping Zhang, Fumiaki Hayashi, Hirokazu Kawagishi, Elyza Aiman Azizah Nur, Hiroshi Tomoda, Elly Wahyudin, Taichi Ohshiro, Jing Wu, and Keisuke Kobayashi
- Subjects
Clinical Biochemistry ,Sterol O-acyltransferase ,Pharmaceutical Science ,CHO Cells ,01 natural sciences ,Biochemistry ,Lactones ,chemistry.chemical_compound ,Cricetulus ,Microsomes ,Drug Discovery ,Animals ,Enzyme Inhibitors ,Cytotoxicity ,Molecular Biology ,Enzyme Assays ,chemistry.chemical_classification ,biology ,010405 organic chemistry ,Chinese hamster ovary cell ,Organic Chemistry ,Polyynes ,Atractylodes ,biology.organism_classification ,Sterol ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,Enzyme ,chemistry ,Cholesteryl ester ,Microsome ,Molecular Medicine ,Cholesterol Esters ,Sesquiterpenes ,Rhizome ,Sterol O-Acyltransferase - Abstract
Using activity guided purification, four known compounds, sesquiterpene atractylenolide III (1), and the polyacetylenes 14-acetoxy-12-senecioyloxytetradeca-2E,8E,10E-trien-4,6-diyn-1-ol (2), 14-acetoxy-12-α-methylbutyl-2E,8E,10E-trien-4,6-diyn-1-ol (3), and 14-acetoxy-12-β -methylbutyl-2E,8E,10E-trien-4,6-diyn-1-ol (4), were isolated from a traditional herbal medicine, Atractylodes rhizome. Structurally similar 3 and 4 (3/4 mixture) were obtained as a mixture. In intact Chinese hamster ovary (CHO) K1 cell assays, 1, 2, and a 3/4 mixture selectively inhibited cholesterol [14C]oleate synthesis from [14C]oleate with IC50 values of 73.5 µM, 35.4 µM, and 10.2 µM, respectively, without any effects on cytotoxicity. As a potential target of these inhibitors involved in cholesteryl ester (CE) synthesis, effects on sterol O-acyltransferase (SOAT) activity were investigated using microsomes prepared from CHO-K1 cells as an enzyme source. Hence, these compounds inhibit SOAT activity with IC50 values (211 µM for 1, 29.0 µM for 2, and 11.8 µM for 3/4 mixture) that correlate well with those measured from intact cell assays. Our results strongly suggest that these compounds inhibit CE synthesis by blocking SOAT activity in CHO-K1 cells.
- Published
- 2020
33. Controlling the degradation of an oxidized dextran-based hydrogel independent of the mechanical properties
- Author
-
Fumiaki Hayashi, Suong-Hyu Hyon, Kazuaki Matsumura, Akimasa Matsugami, and Punnida Nonsuwan
- Subjects
Glycidyl methacrylate ,Polymers and Plastics ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Aldehyde ,Biomaterials ,chemistry.chemical_compound ,Materials Chemistry ,chemistry.chemical_classification ,Organic Chemistry ,Periodate ,021001 nanoscience & nanotechnology ,Combinatorial chemistry ,0104 chemical sciences ,Hydrogel ,Dextran ,chemistry ,Drug delivery ,Thiol ,Biodegradation ,Degradation (geology) ,Glutaraldehyde ,0210 nano-technology ,Aldehyde dextran - Abstract
The objective of this study is to control and elucidate the mechanism of molecular degradation in a polysaccharide hydrogel. Glycidyl methacrylate (GMA) immobilized dextran (Dex-GMA) was oxidized by periodate to introduce aldehyde groups (oxidized Dex-GMA). The hydrogel was formed by the addition of dithiothreitol to the oxidized Dex-GMA solution through thiol Michael addition with the preservation of the aldehyde group for degradation points. It was experimentally determined that the degradation of this hydrogel can be controlled by the addition of amino groups and the speed of degradation can be controlled independently of mechanical properties because crosslinking and degradation points are different. In addition, the molecular mechanism of the crosslinking between the thiol and aldehyde groups was found to control the degradation of dextran derivatives. It is expected that these results will be beneficial in the design of polymer materials in which the speed of degradation can be precisely controlled. In addition, the cytotoxicity of oxidized Dex-GMA was approximately 3000 times lower than that of glutaraldehyde. The low cytotoxicity of the aldehyde in oxidized Dex-GMA was the likely reason for the harmless functionalized polysaccharide material. Possible future clinical applications include cell scaffolds in regenerative medicine and carriers for drug delivery systems.
- Published
- 2018
34. Mirilactams C-E, Novel Polycyclic Macrolactams Isolated from Combined-Culture of Actinosynnema mirum NBRC 14064 and Mycolic Acid-Containing Bacterium
- Author
-
Shotaro, Hoshino, Masahiro, Ozeki, Chin Piow, Wong, Huiping, Zhang, Fumiaki, Hayashi, Takayoshi, Awakawa, Hiroyuki, Morita, Hiroyasu, Onaka, and Ikuro, Abe
- Subjects
Actinobacteria ,Mycolic Acids ,Lactams, Macrocyclic ,Actinomycetales ,Molecular Conformation - Abstract
Mycolic acid-containing bacteria (MACB) are known to activate cryptic natural product biosynthesis in co-cultures with actinobacteria. We cultured Actinosynnema mirum NBRC 14064, a producer of the mono-cyclic polyene macrolactam mirilactam A (6), with the MACB Tsukamurella pulmonis TP-B0596. As a result, three novel compounds (mirilactams C-E, 1-3) were produced in the co-culture conditions. Compounds 1-3 were likely derived from 6 by epoxidation and subsequent spontaneous cyclization. The chemical structures and stereochemistries of 1-3 were determined by spectroscopic analyses (NMR and MS), conformational searches in the optimized potentials for liquid simulations-3 (OPLS3) force field, and calculations of electronic circular dichroism (ECD).
- Published
- 2018
35. Conformation and dynamics of soluble repetitive domain elucidates the initial β-sheet formation of spider silk
- Author
-
Ali D. Malay, David L. Kaplan, Keiji Numata, Akimasa Matsugami, Nur Alia Oktaviani, and Fumiaki Hayashi
- Subjects
0301 basic medicine ,Circular dichroism ,Magnetic Resonance Spectroscopy ,Science ,Silk ,Beta sheet ,General Physics and Astronomy ,macromolecular substances ,010402 general chemistry ,01 natural sciences ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Protein structure ,Tensile Strength ,Animals ,Spider silk ,lcsh:Science ,Polyproline helix ,Multidisciplinary ,Chemistry ,Circular Dichroism ,fungi ,technology, industry, and agriculture ,Spiders ,General Chemistry ,equipment and supplies ,Random coil ,0104 chemical sciences ,030104 developmental biology ,SILK ,Helix ,Biophysics ,lcsh:Q ,Protein Conformation, beta-Strand ,Stress, Mechanical - Abstract
The β-sheet is the key structure underlying the excellent mechanical properties of spider silk. However, the comprehensive mechanism underlying β-sheet formation from soluble silk proteins during the transition into insoluble stable fibers has not been elucidated. Notably, the assembly of repetitive domains that dominate the length of the protein chains and structural features within the spun fibers has not been clarified. Here we determine the conformation and dynamics of the soluble precursor of the repetitive domain of spider silk using solution-state NMR, far-UV circular dichroism and vibrational circular dichroism. The soluble repetitive domain contains two major populations: ~65% random coil and ~24% polyproline type II helix (PPII helix). The PPII helix conformation in the glycine-rich region is proposed as a soluble prefibrillar region that subsequently undergoes intramolecular interactions. These findings unravel the mechanism underlying the initial step of β-sheet formation, which is an extremely rapid process during spider silk assembly., β-sheet structure underlies the mechanical properties of spider silk but the mechanism to form β-sheet from soluble silk protein during transition into insoluble fibers has not been elucidated. Here the authors unravel the mechanism of β-sheet formation using NMR and circular dichroism spectroscopy.
- Published
- 2018
36. Hinduchelins A-D, Noncytotoxic Catechol Derivatives from Streptoalloteichus hindustanus
- Author
-
Ikuro Abe, Huiping Zhang, Hitomi Nakamura, Fei He, Joyce Seow Fong Chin, Shotaro Hoshino, Fumiaki Hayashi, and Liang Yang
- Subjects
Magnetic Resonance Spectroscopy ,Stereochemistry ,Catechols ,Pharmaceutical Science ,Phenylalanine ,Tumor cells ,010402 general chemistry ,medicine.disease_cause ,01 natural sciences ,Analytical Chemistry ,chemistry.chemical_compound ,Cell Line, Tumor ,Drug Discovery ,Actinomycetales ,medicine ,Humans ,Threonine ,Pharmacology ,Quantum chemical ,Catechol ,Pyoverdine ,010405 organic chemistry ,Chemistry ,Pseudomonas aeruginosa ,Organic Chemistry ,Streptoalloteichus hindustanus ,0104 chemical sciences ,Complementary and alternative medicine ,A549 Cells ,MCF-7 Cells ,Molecular Medicine ,Oligopeptides - Abstract
Four new catechol derivatives, hinduchelins A–D (1–4), composed of 2,3- dihydroxybenzoic acid, threonine, and decarboxylated phenylalanine, were isolated from Streptoalloteichus hindustanus. Their structures and absolute configurations were elucidated by interpretation of NMR and HRMS data and quantum chemical ECD calculations. The iron-binding properties of the compounds were evaluated by a pyoverdine production assay in Pseudomonas aeruginosa, and compound 4 showed moderate ability to induce pyoverdine production at 50 μM. None of the compounds were cytotoxic toward HL-20, A549, SMMC-7721, MCF-7, and SW-480 tumor cell lines.
- Published
- 2018
37. Protein cytoplasmic delivery using polyampholyte nanoparticles and freeze concentration
- Author
-
Fumiaki Hayashi, Kazuaki Matsumura, Sana Ahmed, and Toshio Nagashima
- Subjects
Cytoplasm ,Magnetic Resonance Spectroscopy ,Materials science ,Cell Survival ,Static Electricity ,Biophysics ,Nanoparticle ,Bioengineering ,Peptide ,Buffers ,Biomaterials ,chemistry.chemical_compound ,Succinylation ,Cryoprotective Agents ,Drug Delivery Systems ,Adsorption ,Freezing ,Static electricity ,Animals ,Organic chemistry ,Particle Size ,Cryopreservation ,chemistry.chemical_classification ,Protein delivery ,Cationic polymerization ,Succinic anhydride ,Freeze concentration ,Serum Albumin, Bovine ,Polyampholytes ,Endocytosis ,chemistry ,Mechanics of Materials ,Ceramics and Composites ,Nanoparticles ,Cattle ,Muramidase ,Nanocarriers ,Hydrophobic and Hydrophilic Interactions - Abstract
A protein delivery method using freeze concentration was presented with a variety of polyampholyte nanocarriers. In order to develop protein nanocarriers, hydrophobically modified polyampholytes were synthesized by the succinylation of ε-poly-L-lysine with dodecyl succinic anhydride and succinic anhydride. The self-assembled polyampholyte aggregated form nanoparticles through intermolecular hydrophobic and electrostatic interactions when dissolved in aqueous media. The cationic and anionic nanoparticles were easily prepared by changing the succinylation ratio. Anionic or cationic proteins were adsorbed on/into the nanoparticles depending on their surface charges. The protein-loaded nanoparticles were stable for at least 7 d. When L929 cells were frozen with the protein-loaded nanoparticles in the presence of a cryoprotectant, the adsorption of the protein-loaded nanoparticles was enhanced and can be explained by the freeze concentration mechanism. After thawing, proteins were internalized into cells via endocytosis. This was the first report that showed that the efficacy of protein delivery was successfully enhanced by the freeze concentration method. This method could be useful for in vitro cytoplasmic protein or peptide delivery to various cells for immunotherapy or phenotype transformations.
- Published
- 2014
38. Dietziamides, novel tetramic acid dimers from Dietzia timorensis MZ-3 with antioxidative activity
- Author
-
Masahiro Okada, Huiping Zhang, Ikuro Abe, Toshiyuki Wakimoto, Shotaro Hoshino, and Fumiaki Hayashi
- Subjects
Genus Dietzia ,Lactams ,Molecular Structure ,Dietzia timorensis ,Chemistry ,Stereochemistry ,DPPH ,Organic Chemistry ,Clinical Biochemistry ,Pharmaceutical Science ,Tetramic acid ,medicine.disease_cause ,Biochemistry ,Antioxidants ,chemistry.chemical_compound ,Actinomycetales ,Drug Discovery ,medicine ,Molecular Medicine ,Dimerization ,Molecular Biology - Abstract
Dietziamides A and B, two novel tetramic acid dimers, were isolated from the rare actinomycetes Dietzia timorensis MZ-3 in the course of our HPLC-diode array screening of our collection of terrestrial actinomycetes. The spectroscopic analysis revealed the chemical structures of the first secondary metabolites characterized in the genus Dietzia. Dietziamides A and B showed moderate DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activities.
- Published
- 2015
39. Long-term cryopreservation of human mesenchymal stem cells using carboxylated poly-l-lysine without the addition of proteins or dimethyl sulfoxide
- Author
-
Suong-Hyu Hyon, Kazuaki Matsumura, Fumiaki Hayashi, and Toshio Nagashima
- Subjects
Succinic Anhydrides ,Materials science ,Cell Survival ,Cellular differentiation ,Biomedical Engineering ,Biophysics ,Bioengineering ,Osteocytes ,Regenerative medicine ,Cryopreservation ,Biomaterials ,chemistry.chemical_compound ,Cryoprotective Agents ,Tissue engineering ,polyampholytes ,stem cells ,Humans ,Polylysine ,Viability assay ,Cell Proliferation ,Cell growth ,Dimethyl sulfoxide ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,Cell biology ,Biochemistry ,chemistry ,tissue engineering - Abstract
Human bone marrow-derived mesenchymal stem cells (hBMSCs) are known for their potential to undergo mesodermal differentiation into many cell types, including osteocytes, adipocytes, and chondrocytes. Therefore, hBMSCs could be used for a variety of regenerative medicine therapies; in fact, hBMC-derived osteocytes have already been used in bone reconstruction. This study discusses the viability and the differentiation properties of hBMSCs that have been cryopreserved in the absence of proteins or dimethyl sulfoxide (DMSO), by using a novel polyampholyte cryoprotectant (CPA). This CPA is based on carboxylated poly-L-lysine (COOH-PLL) and was prepared by a reaction between -poly-L-lysine and succinic anhydride. 1H-NMR and two-dimensional correlation (1H-13C HSQC) spectroscopy revealed that COOH-PLL did not have a special structure in solution. hBMSCs can be cryopreserved for 24 months at −80°C by using a 7.5% (w/w) cryopreserving solution of COOH-PLL which introduces carboxyl groups that result in >90% cell viability after thawing. Furthermore, the cryopreserved hBMSCs fully retained both their proliferative capacity as well as their potential for osteogenic, adipogenic, and chondrogenic differentiation. Confocal laser-scanning microscopy findings showed that the polyampholyte CPA did not penetrate the cell membrane; rather, it attached to the membrane during cryopreservation. These results indicate that the cryoprotective mechanisms of COOH-PLL might differ from those of currently used small-molecule CPAs. These results also suggest that using COOH-PLL as a cryoprotectant for hBMSC preservation can eliminate the use of proteins and DMSO, which would be safer if these cells were used for cell transplantation or regenerative medicine.
- Published
- 2013
40. Self-Assembled Peptide-Based System for Mitochondrial-Targeted Gene Delivery: Functional and Structural Insights
- Author
-
Fumiaki Hayashi, Keiji Numata, Jo-Ann Chuah, and Akimasa Matsugami
- Subjects
0301 basic medicine ,Polymers and Plastics ,Genetic Vectors ,Bioengineering ,Peptide ,02 engineering and technology ,Endosomes ,Gene delivery ,Mitochondrion ,Biology ,Transfection ,Self assembled ,Biomaterials ,03 medical and health sciences ,chemistry.chemical_compound ,Materials Chemistry ,Humans ,Gene ,chemistry.chemical_classification ,Gene carrier ,Gene Transfer Techniques ,Genetic Therapy ,021001 nanoscience & nanotechnology ,Molecular biology ,Cell biology ,Mitochondria ,030104 developmental biology ,chemistry ,0210 nano-technology ,Peptides ,DNA ,Function (biology) - Abstract
Human mitochondrial dysfunction can lead to severe and often deadly diseases, for which there are no known cures. Although the targeted delivery of therapeutic gene to mitochondria is a promising approach to alleviate these disorders, gene carrier systems for the selective delivery of functional DNA into the mitochondria of living mammalian cells are currently unavailable. Here we rationally developed dual-domain peptides containing DNA-condensing/cell-penetrating/endosome-disruptive and mitochondria-targeting sequences. Secondary structures of the dual-domain peptides were analyzed, and variations in the physicochemical properties (stability, size, and ζ potential) of peptide/DNA complexes were studied as a function of peptide-to-DNA ratio and serum addition. An optimized formulation, identified through qualitative and quantitative studies, fulfills the fundamental prerequisites for mitochondria-specific DNA delivery, successfully transfecting a high proportion (82 ± 2%) of mitochondria in a human cell line with concomitant biocompatibility. Nuclear magnetic resonance studies confirmed the effectiveness of our bipartite peptide design with segregated functions: a helical domain necessary for mitochondrial import and an unstructured region for interaction with DNA involving lysine residues. Further analyses revealed that the lysine-specific interaction assisted the self-organization of the peptide and the DNA cargo, leading to a structural arrangement within the formed complex that is crucial for its biological efficiency. Thus the reported gene vector represents a new and reliable tool to uncover the complexity of mitochondrial transfection.
- Published
- 2016
41. Discovery of Key Dioxygenases that Diverged the Paraherquonin and Acetoxydehydroaustin Pathways in Penicillium brasilianum
- Author
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Ikuro Abe, Huiping Zhang, Yudai Matsuda, Takayuki Fujimoto, Takayoshi Awakawa, Yu Nakashima, Taiki Iwabuchi, Takahiro Mori, and Fumiaki Hayashi
- Subjects
Stereochemistry ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Catalysis ,Gene Expression Regulation, Enzymologic ,Dioxygenases ,Fungal Proteins ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Biosynthesis ,Dioxygenase ,Gene Expression Regulation, Fungal ,Gene cluster ,Moiety ,Gene ,Molecular Structure ,010405 organic chemistry ,Terpenes ,Penicillium ,General Chemistry ,Nonheme iron ,0104 chemical sciences ,chemistry ,Fungal strain ,Penicillium brasilianum ,Genome, Fungal - Abstract
Paraherquonin (1), a fungal meroterpenoid produced by Penicillium brasilianum NBRC 6234, possesses a unique, highly congested hexacyclic molecular architecture. Here we identified the biosynthetic gene cluster of 1 (the prh cluster) and elucidated the pathway up to berkeleydione (2), which serves as the key intermediate for the biosynthesis of 1 as well as many other meroterpenoids. Interestingly, the nonheme iron and α-ketoglutarate-dependent dioxygenase PrhA constructs the cycloheptadiene moiety to afford 2 from preaustinoid A1 (6), probably via the homoallyl-homoallyl radical rearrangement. Additionally, another fungal strain, P. brasilianum MG11, which produces acetoxydehydroaustin instead of 1, was found to have a gene cluster nearly identical to the prh cluster. The dioxygenase encoded by the cluster shares 92% sequence identity with PrhA, and also accepts 6 but produces preaustinoid A3 (17) with a spiro-lactone system, generating a diverging point for the two different meroterpenoid pathways in the same species.
- Published
- 2016
42. ChemInform Abstract: Astellifadiene: Structure Determination by NMR Spectroscopy and Crystalline Sponge Method, and Elucidation of Its Biosynthesis
- Author
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Fumiaki Hayashi, Makoto Fujita, Shoukou Lee, Yudai Matsuda, Manabu Hoshino, Huiping Zhang, Masahiro Okada, Ikuro Abe, Takaaki Mitsuhashi, and Takahiro Mori
- Subjects
chemistry.chemical_classification ,biology ,Chemistry ,Stereochemistry ,Sequence (biology) ,General Medicine ,Nuclear magnetic resonance spectroscopy ,biology.organism_classification ,Terpene ,Sponge ,chemistry.chemical_compound ,Hydrocarbon ,Emericella ,Aspergillus oryzae ,Biosynthesis - Abstract
Genome mining of a terpene synthase gene from Emericella variecolor NBRC 32302 and its functional expression in Aspergillus oryzae led to the production of the new sesterterpene hydrocarbon, astellifadiene (1), having a 6-8-6-5-fused ring system. The structure of 1 was initially investigated by extensive NMR analyses, and was further confirmed by the crystalline sponge method, which established the absolute structure of 1 and demonstrated the usefulness of the method in the structure determination of complex hydrocarbon natural products. Furthermore, the biosynthesis of 1 was proposed on the basis of isotope-incorporation experiments performed both in vivo and in vitro. The cyclization of GFPP involves a protonation-initiated second cyclization sequence, 1,2-alkyl migration, and 1,5-hydride shift to generate the novel scaffold of 1.
- Published
- 2016
43. Rationally designed mineralization for selective recovery of the rare earth elements
- Author
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Takamasa Nonaka, Hideki Takagi, Akimasa Matsugami, Takaaki Hatanaka, Fumiaki Hayashi, Takao Tani, and Nobuhiro Ishida
- Subjects
Lanthanide ,Metal ions in aqueous solution ,Science ,General Physics and Astronomy ,Mineralogy ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Article ,Catalysis ,Metal ,chemistry.chemical_compound ,Multidisciplinary ,Aqueous solution ,Extraction (chemistry) ,General Chemistry ,Mineralization (soil science) ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,chemistry ,Chemical engineering ,visual_art ,visual_art.visual_art_medium ,Hydroxide ,0210 nano-technology - Abstract
The increasing demand for rare earth (RE) elements in advanced materials for permanent magnets, rechargeable batteries, catalysts and lamp phosphors necessitates environmentally friendly approaches for their recovery and separation. Here, we propose a mineralization concept for direct extraction of RE ions with Lamp (lanthanide ion mineralization peptide). In aqueous solution containing various metal ions, Lamp promotes the generation of RE hydroxide species with which it binds to form hydrophobic complexes that accumulate spontaneously as insoluble precipitates, even under physiological conditions (pH ∼6.0). This concept for stabilization of an insoluble lanthanide hydroxide complex with an artificial peptide also works in combination with stable scaffolds like synthetic macromolecules and proteins. Our strategy opens the possibility for selective separation of target metal elements from seawater and industrial wastewater under mild conditions without additional energy input., Lanthanide elements are difficult to separate from aqueous solution with low energy input. Here, the authors design a peptide that recognizes and drives the precipitation of an insoluble lanthanide complex under physiological conditions, introducing a biomineralization-based approach for rare earth recovery.
- Published
- 2016
44. Associations Between Autoimmune Thyroid Disease Prognosis and Functional Polymorphisms of Susceptibility Genes, CTLA4, PTPN22, CD40, FCRL3, and ZFAT, Previously Revealed in Genome-wide Association Studies
- Author
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Hiroya Yamada, Kazuya Takemura, Maiko Akahane, Fumiaki Hayashi, Yu Shimizuishi, Naoya Inoue, Mikio Watanabe, Yoh Hidaka, Noriko Yamakawa, and Yoshinori Iwatani
- Subjects
Adult ,Male ,medicine.medical_specialty ,Genotype ,Hashimoto's disease ,Graves' disease ,Immunology ,Genome-wide association study ,Hashimoto Disease ,Severity of Illness Index ,PTPN22 ,Gene Frequency ,Internal medicine ,medicine ,Humans ,Immunology and Allergy ,CTLA-4 Antigen ,Genetic Predisposition to Disease ,CD40 Antigens ,Receptors, Immunologic ,Allele frequency ,Alleles ,Genetic association ,business.industry ,Case-control study ,Protein Tyrosine Phosphatase, Non-Receptor Type 22 ,Middle Aged ,Prognosis ,medicine.disease ,Graves Disease ,Endocrinology ,Case-Control Studies ,Female ,business ,Polymorphism, Restriction Fragment Length ,Genome-Wide Association Study ,Transcription Factors - Abstract
Genome-wide association studies have revealed several susceptibility genes among patients with autoimmune thyroid disease (AITD), including CTLA4, PTPN22, FCRL3, and ZFAT. However, any possible association between these genes and AITD prognosis remains unknown. The objective of this study was to identify associations between polymorphisms of these genes and AITD prognosis. We genotyped functional polymorphisms, including CTLA4 CT60, CTLA4 +49A/G, CTLA4 -1147C/T, CTLA4 -318C/T, PTPN22 -1123C/G, PTPN22 SNP37, CD40 -1C/T, FCRL3 -169C/T, ZFAT Ex9b-SNP10, and ZFAT Ex9b-SNP2, in 197 AITD patients carefully selected from 456 registered AITD patients, and 86 control subjects. The restriction fragment length polymorphism method was used for genotyping. The CD40 -1CC genotype and C allele were significantly more frequent in patients with Graves’ disease (GD) in remission than in those with intractable GD (P = 0.041 and P = 0.031, respectively). The FCRL3 -169TT genotype was significantly less frequent in patients with intractable GD than in those with GD in remission (P = 0.0324). For a ZFAT Ex9b-SNP10 polymorphism, the TT genotype and T allele were significantly more frequent in patients with severe Hashimoto’s disease (HD) than in those with mild HD (P = 0.0029 and P = 0.0049, respectively). For a CTLA4 CT60 polymorphism, the antithyrotropin receptor antibody levels at the onset of GD were significantly higher in those with the GG genotype than in those with other genotypes (P = 0.0117). CD40 and FCRL3 gene polymorphisms were associated with GD intractability, and ZFAT polymorphism was associated with HD severity but not its development.
- Published
- 2012
45. ZF21 Protein, a Regulator of the Disassembly of Focal Adhesions and Cancer Metastasis, Contains a Novel Noncanonical Pleckstrin Homology Domain
- Author
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Tadashi Tomizawa, Takuya Shuo, Motoharu Seiki, Mikako Shirouzu, Naohiko Koshikawa, Seizo Koshiba, Takanori Kigawa, Daisuke Hoshino, Takushi Harada, Fumiaki Hayashi, Naoya Tochio, Noriko Handa, Satoru Watanabe, Shigeyuki Yokoyama, Toshifumi Akizawa, and Makoto Nagano
- Subjects
Amino Acid Motifs ,Integrin ,Biology ,Biochemistry ,Cell Line ,EEA1 ,Focal adhesion ,Mice ,Protein structure ,Cell Movement ,Neoplasms ,Animals ,Humans ,Neoplasm Metastasis ,Phosphorylation ,Molecular Biology ,Focal Adhesions ,Calpain ,Integrin beta1 ,Intracellular Signaling Peptides and Proteins ,Membrane Proteins ,Molecular Bases of Disease ,Cell migration ,Cell Biology ,Cell biology ,Pleckstrin homology domain ,Focal Adhesion Kinase 1 ,FYVE domain ,biology.protein ,Protein Tyrosine Phosphatases ,Carrier Proteins ,Protein A - Abstract
Directional migration of adherent cells on an extracellular matrix requires repeated formation and disassembly of focal adhesions (FAs). Directional migration of adherent cells We have identified ZF21 as a regulator of disassembly of FAs and cell migration, and increased expression of the gene has been linked to metastatic colon cancer. ZF21 is a member of a protein family characterized by the presence of the FYVE domain, which is conserved among Fab1p, YOPB, Vps27p, and EEA1 proteins, and has been shown to mediate the binding of such proteins to phosphoinositides in the lipid layers of cell membranes. ZF21 binds multiple factors that promote disassembly of FAs such as FAK, β-tubulin, m-calpain, and SHP-2. ZF21 does not contain any other known protein motifs other than the FYVE domain, but a region of the protein C-terminal to the FYVE domain is sufficient to mediate binding to β-tubulin. In this study, we demonstrate that the C-terminal region is important for the ability of ZF21 to induce disassembly of FAs and cell migration, and to promote an early step of experimental metastasis to the lung in mice. In light of the importance of the C-terminal region, we analyzed its ternary structure using NMR spectroscopy. We demonstrate that this region exhibits a structure similar to that of a canonical pleckstrin homology domain, but that it lacks a positively charged interface to bind phosphatidylinositol phosphate. Thus, ZF21 contains a novel noncanonical PH-like domain that is a possible target to develop a therapeutic strategy to treat metastatic cancer.
- Published
- 2011
46. Brevisulcenal-G, -H, and –I, Polycyclic Ether Marine Toxins from the Dinoflagellate Karenia brevisulcata
- Author
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Patrick T. Holland, Huiping Zhang, Fumiaki Hayashi, Veronica Beuzenberg, D. Tim Harwood, Yuka Hamamoto, Feng Shi, Raku Irie, Masayuki Satake, Yoshiyuki Itoh, and Kazuo Tachibana
- Subjects
0301 basic medicine ,Pharmacology ,Karenia brevisulcata ,biology ,Organic Chemistry ,Dinoflagellate ,Ether ,biology.organism_classification ,Analytical Chemistry ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,chemistry ,Environmental chemistry ,Marine toxin - Published
- 2018
47. A B-Box 2 Surface Patch Important for TRIM5α Self-Association, Capsid Binding Avidity, and Retrovirus Restriction
- Author
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Xu Rong Qin, Felipe Diaz-Griffero, Takanori Kigawa, Maritza Lienlaf, Fumiaki Hayashi, Shigeyuki Yokoyama, Zoe Sarnak, Andrés Finzi, and Joseph Sodroski
- Subjects
Models, Molecular ,Magnetic Resonance Spectroscopy ,Ubiquitin-Protein Ligases ,Immunology ,medicine.disease_cause ,Microbiology ,Cell Line ,Capsid ,Retrovirus ,Protein structure ,Virology ,medicine ,Animals ,Humans ,Avidity ,Mutation ,biology ,Structure and Assembly ,Proteins ,biology.organism_classification ,Macaca mulatta ,Molecular biology ,Protein Structure, Tertiary ,Cell biology ,Retroviridae ,Cytoplasm ,Insect Science ,HIV-1 ,biology.protein ,Tripartite Motif Proteins ,TRIM5alpha ,Dimerization - Abstract
TRIM5α is a tripartite motif (TRIM) protein that consists of RING, B-box 2, coiled-coil, and B30.2(SPRY) domains. The TRIM5α rh protein from rhesus monkeys recognizes the human immunodeficiency virus type 1 (HIV-1) capsid as it enters the host cell and blocks virus infection prior to reverse transcription. HIV-1-restricting ability can be eliminated by disruption of the B-box 2 domain. Changes in the TRIM5α rh B-box 2 domain have been associated with alterations in TRIM5α rh turnover, the formation of cytoplasmic bodies and higher-order oligomerization. We present here the nuclear magnetic resonance structure of the TRIM5 B-box 2 domain and identify an unusual hydrophobic patch (cluster 1) on the domain surface. Alteration of cluster 1 or the flanking arginine 121 resulted in various degrees of inactivation of HIV-1 restriction, in some cases depending on compensatory changes in other nearby charged residues. For this panel of TRIM5α rh B-box 2 mutants, inhibition of HIV-1 infection was strongly correlated with higher-order self-association and binding affinity for capsid complexes but not with TRIM5α rh half-life or the formation of cytoplasmic bodies. Thus, promoting cooperative TRIM5α rh interactions with the HIV-1 capsid represents a major mechanism whereby the B-box 2 domain potentiates HIV-1 restriction.
- Published
- 2009
48. Structure of the C-terminal Phosphotyrosine Interaction Domain of Fe65L1 Complexed with the Cytoplasmic Tail of Amyloid Precursor Protein Reveals a Novel Peptide Binding Mode
- Author
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Naoya Tochio, Tadashi Tomizawa, Fumiaki Hayashi, Takuma Kasai, Shigeyuki Yokoyama, Hua Li, Takushi Harada, Akiko Tanaka, Satoru Watanabe, Seizo Koshiba, Yoko Motoda, Takanori Kigawa, Yoshihide Hayashizaki, Takashi Yabuki, and Makoto Inoue
- Subjects
Phosphotyrosine binding ,Protein Folding ,Amino Acid Motifs ,Nerve Tissue Proteins ,Peptide ,Peptide binding ,Biology ,Biochemistry ,Amyloid beta-Protein Precursor ,Mice ,Amyloid precursor protein ,Animals ,Protein Structure, Quaternary ,Nuclear Magnetic Resonance, Biomolecular ,Molecular Biology ,chemistry.chemical_classification ,Nuclear Proteins ,Signal transducing adaptor protein ,Isothermal titration calorimetry ,Cell Biology ,Protein Structure, Tertiary ,chemistry ,biology.protein ,Biophysics ,Protein folding ,Peptides ,Protein Binding ,Binding domain - Abstract
Fe65L1, a member of the Fe65 family, is an adaptor protein that interacts with the cytoplasmic domain of Alzheimer amyloid precursor protein (APP) through its C-terminal phosphotyrosine interaction/phosphotyrosine binding (PID/PTB) domain. In the present study, the solution structures of the C-terminal PID domain of mouse Fe65L1, alone and in complex with a 32-mer peptide (DAAVTPEERHLSKMQQNGYENPTYKFFEQMQN) derived from the cytoplasmic domain of APP, were determined using NMR spectroscopy. The C-terminal PID domain of Fe65L1 alone exhibits a canonical PID/PTB fold, whereas the complex structure reveals a novel mode of peptide binding. In the complex structure, the NPTY motif forms a type-I beta-turn, and the residues immediately N-terminal to the NPTY motif form an antiparallel beta-sheet with the beta5 strand of the PID domain, the binding mode typically observed in the PID/PTB.peptide complex. On the other hand, the N-terminal region of the peptide forms a 2.5-turn alpha-helix and interacts extensively with the C-terminal alpha-helix and the peripheral regions of the PID domain, representing a novel mode of peptide binding that has not been reported previously for the PID/PTB.peptide complex. The indispensability of the N-terminal region of the peptide for the high affinity of the PID-peptide interaction is consistent with NMR titration and isothermal calorimetry data. The extensive binding features of the PID domain of Fe65L1 with the cytoplasmic domain of APP provide a framework for further understanding of the function, trafficking, and processing of APP modulated by adapter proteins.
- Published
- 2008
49. Solution structure of the general transcription factor 2I domain in mouse TFII-I protein
- Author
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Mayumi Yoshida, Makoto Inoue, Masaaki Aoki, Eiko Seki, Takaho Terada, Shigeyuki Yokoyama, Hiroshi Hirota, Takayoshi Matsuda, Y. Doi-Katayama, Yoshihide Hayashizaki, Mikako Shirouzu, Fumiaki Hayashi, Takashi Yabuki, and Takanori Kigawa
- Subjects
Models, Molecular ,Genetics ,General transcription factor ,Stereochemistry ,Molecular Sequence Data ,Protein domain ,DNA-binding domain ,Leucine-rich repeat ,Biology ,Biochemistry ,Pentapeptide repeat ,Protein Structure, Tertiary ,Solutions ,Mice ,Transcription Factors, TFII ,Protein structure ,Protein Structure Report ,Animals ,Amino Acid Sequence ,B3 domain ,Nuclear Magnetic Resonance, Biomolecular ,Sequence Alignment ,Molecular Biology ,Protein secondary structure - Abstract
The general transcription factor TFII-I, with the corresponding gene name GTF2I, is an unusual transcriptional regulator that associates with both basal and signal-induced transcription factors. TFII-I consists of six GTF2I repeat domains, called I-repeats R1-R6. The structure and function of the GTF2I domain are not clearly understood, even though it contains a helix-loop-helix motif, which is considered to be the protein-protein interaction area, based on biochemical analyses. Here, we report the solution structure of the fifth repeat of the six GTF2I repeat domains from murine TFII-I, which was determined by heteronuclear multidimensional NMR spectroscopy (PDB code 1Q60). The three-dimensional structure of the GTF2I domain is classified as a new fold, consisting of four helices (residues 8-24, 34-39, 63-71, and 83-91), two antiparallel beta strands (residues 44-47 and 77-80), and a well-defined loop containing two beta-turns between sheet 1 and helix 3. All of the repeats probably have similar folds to that of repeat 5, because the conserved residues in the GTF2I repeat domains are assembled on the hydrophobic core, turns, and secondary structure elements, as revealed by a comparison of the sequences of the first through the sixth GTF2I repeats in TFII-I.
- Published
- 2007
50. The association between a functional polymorphism in theCD24gene and the development of autoimmune thyroid diseases
- Author
-
Fumiaki Hayashi, Naoya Inoue, Yoshinori Iwatani, Mikio Watanabe, and Yoh Hidaka
- Subjects
business.industry ,CD24 ,Immunology ,Thyroid ,Case-control study ,Autoimmune thyroid disease ,General Medicine ,Biochemistry ,medicine.anatomical_structure ,Genetics ,Immunology and Allergy ,Medicine ,business ,Gene ,Allele frequency ,Functional polymorphism - Published
- 2013
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