研究者詳細 - 安永　卓生

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85133102192&origin=inward

Extended supercomplex contains type-II NADH dehydrogenase, cytochrome bcc complex, and aa<inf>3</inf> oxidase in the respiratory chain of Corynebacterium glutamicum 査読有り国際誌

Takazaki H., Kusumoto T., Ishibashi W., Yasunaga T., Sakamoto J.

Journal of Bioscience and Bioengineering 133 ( 1 ) 76 - 82 2022年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

To clarify the precise subunit composition of the respiratory supercomplex of Corynebacterium glutamicum, several wash conditions were examined. MEGA (9 + 10) wash-buffer (0.5%) was used for this purpose and two-step column chromatography was performed. Almost equal amounts of cytochrome c, b, and a were observed in the purified fraction, estimated by their different absorption spectra. The 833 kDa and 685 kDa bands were observed in the clear native polyacrylamide gel electrophoresis (CN-PAGE) of the purified fraction. Both bands were stained using N,N',N′,N-tetramethyl-p-phenylenediamine (TMPD) oxidase dye, and the 833 kDa band was also stained using NADH oxidase dye. The 3D map reconstructed from the 833 kDa band indicated that the bcc complex and aa3 oxidase are heterodimers. Lastly, electron transfer from NADH to the bcc-aa3 supercomplex was observed. The 833 kDa band is the supercomplex, which includes the heterodimer cytochrome bcc complex and cytochrome aa3 oxidase, as well as the monomer NDH-II. Hence, we termed the 833 kDa band the extended supercomplex (ESC).

DOI： 10.1016/j.jbiosc.2021.10.004

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Filopodia formation by crosslinking of F-actin with fascin in two different binding manners 査読有り国際誌

Shinji Aramaki, Kouta Mayanagi, Mingyue Jin, Kazuhiro Aoyama, Takuo Yasunaga

Cytoskeleton ( Wiley ) 73 ( 7 ) 365 - 374 2016年06月

Evaluation of a 2k CCD camera with an epitaxially grown CsI scintillator for recording energy-filtered electron cryo-micrographs 査読有り

YASUNAGA Takuo, WAKABAYASHI Takeyuki

Journal of electron microscopy 57 ( 3 ) 101 - 112 2008年06月

Atomic model construction of protein complexes from electron micrographs and visualization of their 3D structure using a virtual reality system 査読有り国際誌

Noda K., Nakamura M., Nishida R., Yoneda Y., Yamaguchi Y., Tamura Y., Nakamura H., Yasunaga T.

Journal of Plasma Physics 72 ( 6 ) 1037 - 1040 2006年12月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

The methods and applications that integrate electron microscopic density maps and comparative atomic coordinates to generate atomic models of protein complexes and their assemblies have recently advanced. Here, we also report on a newly developed tool for integration between density maps at a spatial resolution of about one nanometer and atomic coordinates by introducing the spatial constraints into a molecular dynamics calculation, which we call spatially restricted molecular dynamics. We, successfully, constructed the atomic models well-fitted to the density maps generated from nine different atomic coordinates of proteins. The method will give us the useful, refined, and presumable atomic structure of macromolecules to elucidate the relationship between their structure and functions. © 2006 Cambridge University Press.

DOI： 10.1017/S0022377806005174

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=33846100102&origin=inward

計測分析を未来につなぐためのデータの在り方と標準化招待有り

安永卓生

計測標準と計量管理 ( （一社）日本計量振興協会 ) 73 ( 4 ) 4 - 12 2024年02月

高等学校の探究科目／情報科目におけるシミュレーションの位置づけに関する一考査読有り

安永卓生

シミュレーション ( 日本シミュレーション学会 ) 42 ( 3 ) 188 - 193 2023年09月

計測分析機器の出力データフォーマット共通化査読有り

一村信吾, 重藤知夫, 安永卓生, 井上信介

応用物理 92 ( 3 ) 142 - 146 2023年03月

A percentile rank score of group productivity: an evaluation of publication productivity for researchers from various fields 査読有り国際誌

Yamamoto K., Yasunaga T.

Scientometrics 127 ( 4 ) 1737 - 1754 2022年04月

担当区分：最終著者記述言語：英語掲載種別：研究論文（学術雑誌）

The difficulty in evaluating the research performance of groups is attributable to the following two factors: 1) difference of population size or discipline of group members and 2) skewed distribution of the research performance of individuals. This study attempts to overcome this difficulty, focusing on the research performance based on publication productivity. We employ the normalized index for the number of papers, in which publication efficiency was considered and disciplinary variation in the publication intensity was corrected by the disciplinary averages, to calculate a new percentile rank score. The score was developed on the basis of the principle that a person who is rare is valuable. The score was also tested with publication data for faculty members of 17 Japanese universities. The employment of the normalized index increased the score of universities with relatively few faculty members working in the disciplines of high productivity, resulting in more plausible university rankings. The rankings show a high correlation with those for a previously established percentile rank score, which was developed for citation analysis, and they are consistent with the judgment by evaluators of several universities under study. The advantage of the new score over the previous one is that it has no room for arbitrariness in determining the scheme of rank classification and the weights given to each rank class.

DOI： 10.1007/s11192-022-04278-w

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85125519232&origin=inward

巨大システムとしてのクライオ電子顕微鏡法の自動化から自働化への展開「（共著）」招待有り査読有り

安永卓生, 田中康太郎, 岩嵜彩夏, 塚本崇文, 本多康久

顕微鏡 ( 公益社団法人日本顕微鏡学会 ) 56 ( 1 ) 43 - 47 2021年04月

担当区分：筆頭著者記述言語：日本語掲載種別：記事・総説・解説・論説等（学術雑誌）

クライオ電子顕微鏡を利活用する時代が到来した．単粒子解析法は，原子レベルでの構造解析により構造生物学の柱となってきた．また，電子線トモグラフィー法は，細胞や組織における新しい描像を示しつつある．しかしSN比の低い画像であるが故に，多量の画像を必要とする．そのため，適切なUIに加え，自動撮影と画像処理システムと組み合わせ，画像の質を迅速に評価せねばならない．自動化から自働化へ，創発的な振る舞いが次の時代を興す．

DOI： 10.11410/kenbikyo.56.1_43

Kyutacar

その他リンク： http://hdl.handle.net/10228/00008831

九州工業大学における多面的・総合的な入試の制度設計検証――評価結果および「ジェネリック・スキル」等の比較からの一考察―― 査読有り

花堂奈緒子，播磨良輔，安永卓生

大学入試研究ジャーナル 2021年03月

生物物理学を牽引する新技術査読有り

安藤敏夫, 伊藤隆, 杉田有治, 南後恵理子, 安永卓生, 岡田康志, 神田元紀

生物物理 ( 一般社団法人日本生物物理学会 ) 61 ( 2 ) 111 - 118 2021年01月

Conformational equilibrium of NADPH–cytochrome P450 oxidoreductase is essential for heme oxygenase reaction 査読有り

Sugishima M., Taira J., Sagara T., Nakao R., Sato H., Noguchi M., Fukuyama K., Yamamoto K., Yasunaga T., Sakamoto H.

Antioxidants 9 ( 8 ) 1 - 13 2020年08月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Heme oxygenase (HO) catalyzes heme degradation using electrons supplied by NADPH–cytochrome P450 oxidoreductase (CPR). Electrons from NADPH flow first to FAD, then to FMN, and finally to the heme in the redox partner. Previous biophysical analyses suggest the presence of a dynamic equilibrium between the open and the closed forms of CPR. We previously demonstrated that the open-form stabilized CPR (∆TGEE) is tightly bound to heme–HO-1, whereas the reduction in heme–HO-1 coupled with ∆TGEE is considerably slow because the distance between FAD and FMN in ∆TGEE is inappropriate for electron transfer from FAD to FMN. Here, we characterized the enzymatic activity and the reduction kinetics of HO-1 using the closed-form stabilized CPR (147CC514). Additionally, we analyzed the interaction between 147CC514 and heme–HO-1 by analytical ultracentrifugation. The results indicate that the interaction between 147CC514 and heme–HO-1 is considerably weak, and the enzymatic activity of 147CC514 is markedly weaker than that of CPR. Further, using cryo-electron microscopy, we confirmed that the crystal structure of ∆TGEE in complex with heme–HO-1 is similar to the relatively low-resolution structure of CPR complexed with heme–HO-1 in solution. We conclude that the “open–close” transition of CPR is indispensable for electron transfer from CPR to heme–HO-1.

DOI： 10.3390/antiox9080673

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85088977608&origin=inward

入学者選抜と各種評価法による伸びしろの分析査読有り

安永卓生，播磨良輔，藤江美奈，山下修充

大学入試研究ジャーナル 2020年03月

Molecular dynamics study on DNA damage by tritium disintegration 査読有り

Nakamura H., Miyanishi H., Yasunaga T., Fujiwara S., Mizuguchi T., Nakata A., Miyazaki T., Otsuka T., Kenmotsu T., Hatano Y., Saito S.

Japanese Journal of Applied Physics 59 ( SA ) 2020年01月

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）

© 2019 The Japan Society of Applied Physics. Using molecular dynamics (MD) simulation, we simulate the structural change of a telomeric DNA by β-decay of substituted tritium to helium-3. The configuration of the telomeric DNA is obtained by removing TRF2 protein from the TRF2-Dbd-DNA complex (Protein Data Bank ID is 3SJM). We assume that hydrogens (H) of guanines in the telomeric DNA are replaced to helium-3. Since this replacement of the H atoms to the 3He atoms changes the charge distribution significantly, the charge distribution used in the MD simulation for the modified guanine is obtained by the density functional theory calculations. We adopt, as the MD simulation, nanoscale molecular dynamics code with CHARMM36 force field using Langevin thermostat and Nosé-Hoover Langevin piston to control the temperature and pressure of the system, respectively. Moreover, changing both the number of replaced guanine N and the temperature of the system T, we calculate the root mean square deviation RMSD to quantify the dependence of the durability of the telomeric DNA on the β-decays. From the MD simulation, it is found that as N or T becomes larger, the RMSD of the DNA becomes also larger. Namely, it denotes that as the intensity of the β-decays becomes larger or as the temperature is increased, the DNA structure becomes more fragile.

DOI： 10.7567/1347-4065/ab460d

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85079777993&origin=inward

顕微鏡学における新たな鏡としての機械学習招待有り査読有り

安永卓生

顕微鏡 ( 公益社団法人日本顕微鏡学会 ) 55 ( 3 ) 103 - 103 2020年01月

難加工研磨におけるフラーレン結合型機能性ナノ微粒子に関する研究査読有り

久吉楓也, 鈴木恵友, Tsai Yueh-Hsun, Khajornrungruang Panart, 安永卓生, 高崎寛子, 大内将吉

精密工学会学術講演会講演論文集 ( 公益社団法人精密工学会 ) 2020 ( 0 ) 486 - 487 2020年01月

記述言語：日本語掲載種別：研究論文（研究会，シンポジウム資料等）

本研究では粒径が数十nm〜百nm程度の粒子径を持つダイヤモンドなどコア粒子上にフラーレン分子を吸着させ，紫外線照射のほかに，マイクロ波合成法を併用させることでフラーレンを新規炭素同素体へ物質変換を行い，研磨微粒子上に安定化した数nmレベルの機能性炭素層を形成する．ここでは形成された新規炭素同素体の分類を行うとともに難加工基板に対する研磨性能の影響について評価する．

DOI： 10.11522/pscjspe.2020S.0_486

その他リンク： https://ci.nii.ac.jp/naid/130007896718

単粒子解析におけるタンパク質構造分類のための深層学習アプローチの動向招待有り査読有り

馬水信弥, 田中康太郎, 安永卓生

顕微鏡 ( 公益社団法人日本顕微鏡学会 ) 55 ( 3 ) 104 - 108 2020年01月

担当区分：責任著者記述言語：日本語掲載種別：記事・総説・解説・論説等（学術雑誌）

クライオ電子顕微鏡による単粒子解析では，試料中に含まれる複数のタンパク質構造を分類しながら解くことが出来る．ただし分類された構造間のダイナミクスの情報は類推するしかない．この問題について2020年に発表された三次元再構成およびクラス分類を行うための深層学習アプローチであるcryoDRGNは，離散的なデータ分割による構造分類を脱却し，連続的な構造分類を実現した．そこでは，オートエンコーダーをベースとし，入力粒子画像から投影パラメーターに依存する情報を分離して潜在空間を構築している．本稿では従来の構造分類と，cryoDRGNおよびその背景となる深層学習のトピックについて解説を行ったのち，構造分類のベンチマークとして6種類の複合体を有するGroEL/ESの実データについて三次元再構成とその分類を試みた．

DOI： 10.11410/kenbikyo.55.3_104

Kyutacar

その他リンク： https://ci.nii.ac.jp/naid/130007967723

Molecular dynamics study on DNA damage by tritium disintegration 査読有り

Hiroaki Nakamura1,2, Hisanori Miyanishi2, Takuo Yasunaga3, Susumu Fujiwara4, Tomoko Mizuguchi4, Ayako Nakata5, Tsuyoshi Miyazaki5, Takao Otsuka6, Takahiro Kenmotsu7, Yuji Hatano8 and Shinji Saito

Japanese Journal of Applied Physics 59 2019年11月

Structural Changes in Tritium-Substituted Polymeric Materials by Beta Decays: A Molecular Dynamics Study 査読有り国際誌

LI Haolun, FUJIWARA Susumu, NAKAMURA Hiroaki, MIZUGUCHI Tomoko, YASUNAGA Takuo, OTSUKA Takao, KENMOTSU Takahiro, HATANO Yuji, SAITO Shinji

Plasma and Fusion Research ( 一般社団法人プラズマ・核融合学会 ) 14 ( 0 ) 3401106 - 3401106 2019年08月

記述言語：英語掲載種別：研究論文（学術雑誌）

The molecular mechanism through which how beta decays in tritium-substituted species damage DNA and polymeric materials is still unknown. Molecular dynamics simulations of hydrogen-removed polyethylene were performed to predict the structural change of the polyethylene chain after the substituted tritium decays. We calculated the potential energy, the global orientational order parameter, and the average number of consecutive trans bonds. The results are that, the greater the number of removed hydrogen atoms, the higher the potential energy and the lower the value of the global orientational order parameter and the average number of consecutive trans bonds. Thus, after losing hydrogen, polyethylene becomes poorer in terms of both thermal and structural stabilities.

DOI： 10.1585/pfr.14.3401106

その他リンク： https://nifs-repository.repo.nii.ac.jp/records/11296

九州工業大学における入試区分毎の PROG・GPA を用いた追跡調査と制度設計査読有り

安永卓生，藤江美奈，山本鉱，播磨良輔，山下修充

大学入試研究ジャーナル 2019年03月

水和したままのタンパク質や細胞を観る招待有り査読有り

安永卓生

日本結晶学会誌 ( 日本結晶学会 ) 61 ( 1 ) 43 - 50 2019年02月

担当区分：筆頭著者,　最終著者,　責任著者記述言語：日本語掲載種別：記事・総説・解説・論説等（学術雑誌）

The electron cryo-microscopy is one of the most powerful techniques to elucidate biological molecular structure under nearly physiological condition. By single particle analysis, many three-dimensional structures of proteins and their complexes have been resolved at atomic resolutions. Furthermore, in situ molecular behaviours in cells and tissues have also been reported at nano-meter or sub-nanometer resolutions by electron tomography. I would here review current situations of electron cryo-microscopy and the principles of their analysis and so show its possibility.

DOI： 10.5940/jcrsj.61.43

その他リンク： https://www.jstage.jst.go.jp/article/jcrsj/61/1/61_43/_pdf

Computational strategy for studying structural change of tritium-substituted macromolecules by a beta decay to helium-3 査読有り国際誌

Fujiwara Susumu, Nakamura Hiroaki, Li Haolun, Miyanishi Hisanori, Mizuguchi Tomoko, Yasunaga Takuo, Otsuka Takao, Hatano Yuji, Saito Shinji

日本シミュレーション学会英文誌 ( 一般社団法人日本シミュレーション学会 ) 6 ( 1 ) 94 - 99 2019年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

We propose a computational strategy for investigating structural change of tritium-substituted macromolecules. Effects of radiation on macromolecules such as polymeric materials and DNA are classified into three categories: (1) direct action, (2) indirect action, and (3) decay effect. In this study, we focus on the decay effect exclusively. After a beta decay of substituted tritium in macromolecules to helium-3, the generated inert helium-3 is assumed to be deleted quickly. To get an insight into the decay effect to the damage of macromolecules, we perform molecular dynamics simulations of tritium-deleted macromolecules and analyze their structural change. Preliminary simulation results of decay effect on polymeric materials and DNA are presented.

DOI： 10.15748/jasse.6.94

その他リンク： https://www.jstage.jst.go.jp/article/jasse/6/1/6_94/_pdf

3次元クライオ電子顕微鏡法が解き明かす細胞の運動招待有り査読有り

安永卓生, 荒牧慎二, 肥後智也

顕微鏡 ( 公益社団法人日本顕微鏡学会 ) 53 ( 3 ) 118 - 122 2018年12月

担当区分：筆頭著者,　責任著者記述言語：日本語掲載種別：記事・総説・解説・論説等（学術雑誌）

クライオ電子顕微鏡法は，生理的条件に近い状態で分子構造を明らかにできる強力な手法の1つである．電子線トモグラフィー法と組み合わせることで，細胞や組織内での分子動態をナノレベルの分解能で明らかにすることができる．我々は，細胞運動や細胞センシングにとって重要な構造である糸状仮足のアーキテクチャを明らかにしてきた．力学的バランスから推定される本数のアクチン繊維束のセットが観測され，架橋構造とX線結晶解析法による原子モデルと統合し，束化因子ファシンのアクチンへの結合様式を示した．また，アクチン束周辺のファシンの協働的結合や短いアクチン等のブラウジングの結果として，アクチン細胞骨格の束化メカニズムを提唱した．さらに，糸状仮足先端でのアクチン繊維の振る舞いも明らかにしている．今後，クライオ電子線トモグラフィー法は，生物試料のみならず，燃料電池を始めとする水系の材料分野でも有効であろう．

DOI： 10.11410/kenbikyo.53.3_118

その他リンク： https://search.jamas.or.jp/link/ui/2019210169

Correction: Alpha-synuclein facilitates to form short unconventional microtubules that have a unique function in the axonal transport (Scientific Reports DOI: 10.1038/s41598-017-15575-3) 査読有り

Toba S., Jin M., Yamada M., Kumamoto K., Matsumoto S., Yasunaga T., Fukunaga Y., Miyazawa A., Fujita S., Itoh K., Fushiki S., Kojima H., Wanibuchi H., Arai Y., Nagai T., Hirotsune S.

Scientific Reports ( Scientific Reports ) 8 ( 1 ) 2018年12月

掲載種別：記事・総説・解説・論説等（大学・研究所紀要）

© 2018 The Author(s). In this Article, Figure 5 was inadvertently published as Figures 5 and 6, leading to the incorrect publication of Figure 6 as Figure 7 and the omission of the correct Figure 7. The correct Figures 5, 6, and 7 appear below as Figures 1, 2, and 3 respectively. The Figure legends are correct. (Figure Presented).

DOI： 10.1038/s41598-018-25979-4

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85047096548&origin=inward

高大接続改革と九州工大

安永卓生

明専会報 ( 明専会 ) 2018年07月

Effects of mixed ultrafine colloidal silica particles on chemical mechanical polishing of sapphire 査読有り

Bun-Athuek Natthaphon, Takazaki Hiroko, Yoshimoto Yutaka, Khajornrungruang Panart, Yasunaga Takuo, Suzuki Keisuke

Jpn. J. Appl. Phys. 57 ( 7 ) 2018年06月

記述言語：英語掲載種別：研究論文（学術雑誌）

The effects of ultrafine colloidal silica particles adsorbed on the surfaces of large silica particles in slurries used for the chemical mechanical polishing of sapphire were studied. Sapphire wafers were polished using hybrid silica particles that were composed of a mixture of ultrafine (4 nm) colloidal silica particles and large (20, 55, and 105 nm) silica particles. Dynamic light scattering results showed that the hybrid particles became larger after mixing. Transmission electron microscopy images revealed that the ultrafine particles adhered to the surfaces of the large particles and that the surface shape of the hybrid particles were changed by the mixing process. Atomic force microscopy and polishing results showed that the surface roughnesses and material removal rates of sapphire substrates were improved by using the hybrid particles. The coefficient of friction between the surface of the sapphire substrate and the hybrid particles was higher than that obtained using single-sized particles. These results confirm that the hybrid silica particles enhanced the performance of sapphire polishing.

DOI： 10.7567/JJAP.57.07MD03

その他リンク： https://ci.nii.ac.jp/naid/150000116077

生命を解き明かすクライオ電子顕微鏡法の新時代招待有り査読有り

安永卓生

顕微鏡 ( 公益社団法人日本顕微鏡学会 ) 53 ( 1 ) 2 2018年04月

クライオ電子顕微鏡法を支える画像解析と三次元像構築ーその原理と最近のソフトウェアの発展

安永卓生, 荒牧慎二

実験医学 ( 羊土社 ) 36 ( 8 ) 2018年04月

生命を解き明かすクライオ電子顕微鏡法の新時代

安永卓生

顕微鏡 ( 日本顕微鏡学会 ) 1 ( 53 ) 2018年04月

高大接続改革と九州工大 ―学生とのよりよいマッチングを求めて　類別入試と学部・学科の改組 ―

安永卓生

明専会報 ( 明専会 ) 2018年01月

難加工材料におけるハイブリッド研磨微粒子の最適構造に関する研究:化学的作用と機械的作用に関する考察査読有り

鈴木恵友, Bun-Athuek Natthaphon, 髙﨑寛子, 安永卓生, 吉本裕, パナートカチョーンルンルアン

精密工学会学術講演会講演論文集 ( 公益社団法人精密工学会 ) 2018 ( 0 ) 312 - 313 2018年01月

記述言語：日本語掲載種別：研究論文（学術雑誌）

本研究ではコア微粒子上に直径4nmのナノシリカ微粒子や水酸化フラーレンを吸着により生成したハイブリット研磨微粒子を難加工研磨技術への適用を試みた．ここではコアシリカ微粒子の粒子径やナノシリカ微粒子水酸化フラーレンの吸着状態によりサファイアCMPの材料除去レートが変化した．そのためハイブリット研磨微粒子の構造を最適化することにより，化学的作用や機械的作用を独立に制御する手法を試みたので報告する．

DOI： 10.11522/pscjspe.2018A.0_312

その他リンク： https://ci.nii.ac.jp/naid/130007602798

Alpha-synuclein facilitates to form short unconventional microtubules that have a unique function in the axonal transport 査読有り国際誌

Toba S., Jin M., Yamada M., Kumamoto K., Matsumoto S., Yasunaga T., Fukunaga Y., Miyazawa A., Fujita S., Itoh K., Fushiki S., Kojima H., Wanibuchi H., Arai Y., Nagai T., Hirotsune S.

Scientific Reports 7 ( 1 ) 2017年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2017 The Author(s). Although α-synuclein (αSyn) has been linked to Parkinson's disease (PD), the mechanisms underlying the causative role in PD remain unclear. We previously proposed a model for a transportable microtubule (tMT), in which dynein is anchored to a short tMT by LIS1 followed by the kinesin-dependent anterograde transport; however the mechanisms that produce tMTs have not been determined. Our in vitro investigations of microtubule (MT) dynamics revealed that αSyn facilitates the formation of short MTs and preferentially binds to MTs carrying 14 protofilaments (pfs). Live-cell imaging showed that αSyn co-transported with dynein and mobile βIII-tubulin fragments in the anterograde transport. Furthermore, bi-directional axonal transports are severely affected in αSyn and γSyn depleted dorsal root ganglion neurons. SR-PALM analyses further revealed the fibrous co-localization of αSyn, dynein and βIII-tubulin in axons. More importantly, 14-pfs MTs have been found in rat femoral nerve tissue, and they increased approximately 19 fold the control in quantify upon nerve ligation, indicating the unconventional MTs are mobile. Our findings indicate that αSyn facilitates to form short, mobile tMTs that play an important role in the axonal transport. This unexpected and intriguing discovery related to axonal transport provides new insight on the pathogenesis of PD.

DOI： 10.1038/s41598-017-15575-3

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85035338808&origin=inward

Katanin p80, NuMA and cytoplasmic dynein cooperate to control microtubule dynamics 査読有り

Jin M., Pomp O., Shinoda T., Toba S., Torisawa T., Furuta K., Oiwa K., Yasunaga T., Kitagawa D., Matsumura S., Miyata T., Tan T., Reversade B., Hirotsune S.

Scientific Reports 7 2017年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

© The Author(s) 2017. Human mutations in KATNB1 (p80) cause severe congenital cortical malformations, which encompass the clinical features of both microcephaly and lissencephaly. Although p80 plays critical roles during brain development, the underlying mechanisms remain predominately unknown. Here, we demonstrate that p80 regulates microtubule (MT) remodeling in combination with NuMA (nuclear mitotic apparatus protein) and cytoplasmic dynein. We show that p80 shuttles between the nucleus and spindle pole in synchrony with the cell cycle. Interestingly, this striking feature is shared with NuMA. Importantly, p80 is essential for aster formation and maintenance in vitro. siRNA-mediated depletion of p80 and/or NuMA induced abnormal mitotic phenotypes in cultured mouse embryonic fibroblasts and aberrant neurogenesis and neuronal migration in the mouse embryonic brain. Importantly, these results were confirmed in p80-mutant harboring patient-derived induced pluripotent stem cells and brain organoids. Taken together, our findings provide valuable insights into the pathogenesis of severe microlissencephaly, in which p80 and NuMA delineate a common pathway for neurogenesis and neuronal migration via MT organization at the centrosome/spindle pole.

DOI： 10.1038/srep39902

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85009442774&origin=inward

Lis1 restricts the conformational changes in cytoplasmic dynein on microtubules（共著）査読有り

S Toba, K Koyasako, T Yasunaga, S Hirotsune

Microscopy 64 ( 6 ) 419 - 427 2015年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Cytoplasmic dynein is a microtubule-based motor protein that transports intracellular cargo and performs various functions during cell division. We previously reported that Lis1 suppressed dynein motility on microtubules in an idling state. Recently, a model showed that Lis1 prevents the ATPase domain of dynein from transmitting a detachment signal to its micro- tubule-binding domain. However, conformational information on dynein is limited. We used electron microscopy to investigate the conformation of dynein and nucleotide-induced conformational changes on microtubules. The conformation of dynein differed depending on the presence or absence of a nucleotide. In the presence of the nucleotide ADP-vanadate, dynein displayed an extended form on microtubules (extended form), whereas in the absence of a nucleotide, dynein lay along microtubules (compact form). This conformational change reflects chemomechanical coupling in dynein walking on microtubules. We also found that Lis1 fixed the conformation of dynein in the compact form regardless of the nucleotide condition. Removal of the Lis1 dimerization motif abolished Lis1-dependent fixation of dynein in the compact form. This suggests that the idling state of dynein on microtubules induced by Lis1 occurs through the Lis1-dependent arrest of dynein chemomechanical coupling.

DOI： 10.1093/jmicro/dfv055

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84952772401&origin=inward

問題を解決し続ける生物と工学への玄妙の関

安永卓生

生物物理 ( 生物物理学会 ) 2015年09月

GM130 is a parallel tetramer with a flexible rod-like structure and N-terminally open (Y-shaped) and closed (I-shaped) conformations 査読有り

Ryuichi Ishida, Akitsugu Yamamoto, Kazuhisa Nakayama, Miwa Sohda, Yoshio Misumi, Takuo Yasunaga, Nobuhiro Nakamura

FEBS Journal ( Wiley ) 282 ( 11 ) 2232 - 2244 2015年04月

問題を解決し続ける生物と工学への玄妙の関招待有り査読有り

安永卓生

生物物理 ( 一般社団法人日本生物物理学会 ) 55 ( 5 ) 231 - 231 2015年01月

Transformation from Multi- to Single-lamellar Vesicle by Addition of a Cationic Lipid to 1,2-Dilauroyl-sn-glycero-3-phosphocholine Explored with SAXS and TEM 査読有り国際誌

Kubo Genki, Sakamoto Shunsuke, Fujii Shota, Sanada Yusuke, Yasunaga Takuo, Takahara Atsushi, Sakurai Kazuo

Chemistry Letters ( 公益社団法人日本化学会 ) 43 ( 11 ) 1785 - 1787 2014年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

1,2-Dilauroyl-sn-glycero-3-phosphocholine (DLPC) is one of the major lipids constituting cellular membranes. When DLPC itself is dispersed in aqueous solutions, multilamellar vesicles are formed, although the cellular membranes are made of single bilayers. We found that the addition of a calix[4]arene derivative (CaL[4]C3) bearing four amine groups as the headgroup to the DLPC multilamellar vesicle induced the structural transformation to a single-lamellar vesicle, by use of small-angle X-ray scattering and electron microscopy, indicating that the highly condensed ionic groups of CaL[4]C3 are useful to control membrane morphology.

DOI： 10.1246/cl.140668

その他リンク： http://www.journal.csj.jp/doi/pdf/10.1246/cl.140668

1P192 微小管結合タンパク質シヌクレインは細胞質ダイニンを細胞辺縁部に運ぶ機能を持つ輸送性微小管の形成に必須である(12. 細胞生物的課題,ポスター,第52回日本生物物理学会年会(2014年度))

Toba Shiori, Koyasako Kotaro, Yamada Masami, Yasunaga Takuo, Kojima Hiroaki, Wanibuchi Hideki, Hirotsune Shinji

生物物理 ( 一般社団法人日本生物物理学会 ) 54 ( 1 ) S172 2014年01月

1P295 Metallothionein labeling for CLEM(Correlative Light and Electron Microscopy) method(27. Bioimaging,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014)) 査読有り

Yamanaka Ryutaro, Hirasaka Yuka, Jin Mingyue, Yanagisawa Haruaki, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 54 ( 1 ) S190 2014年01月

3P294 短粒子解析法の高速化のためのGPGPUを用いた並列化処理の実装と評価(27. バイオイメージング,ポスター,第52回日本生物物理学会年会(2014年度)) 査読有り

Iwasaki Ayaka, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 54 ( 1 ) S297 2014年01月

Primary cilia-mediated cellular signaling in dermal papilla cells 査読有り

K Matsushima, M Suematsu, C Mifude, S Aramaki, M Tanaka, T Yasunaga, K Kaseda

JOURNAL OF INVESTIGATIVE DERMATOLOGY 133 ( 5 ) 1407 - 1407 2013年05月

Kinesin-1 regulates dendrite microtubule polarity in Caenorhabditis elegans 査読有り国際誌

Jing Yan, Dan L Chao, Shiori Toba, Kotaro Koyasako, Takuo Yasunaga, Shinji Hirotsune, Kang Shen

eLife ( eLife Sciences Publications, Ltd ) 2 e00133 2013年03月

記述言語：英語掲載種別：研究論文（学術雑誌）

<jats:p>In neurons, microtubules (MTs) span the length of both axons and dendrites, and the molecular motors use these intracellular ‘highways' to transport diverse cargo to the appropriate subcellular locations. Whereas axonal MTs are organized such that the plus-end is oriented out from the cell body, dendrites exhibit a mixed MTs polarity containing both minus-end-out and plus-end-out MTs. The molecular mechanisms underlying this differential organization, as well as its functional significance, are unknown. Here, we show that kinesin-1 is critical in establishing the characteristic minus-end-out MT organization of the dendrite in vivo. In unc-116 (kinesin-1/kinesin heavy chain) mutants, the dendritic MTs adopt an axonal-like plus-end-out organization. Kinesin-1 protein is able to cross-link anti-paralleled MTs in vitro. We propose that kinesin-1 regulates the dendrite MT polarity through directly gliding the plus-end-out MTs out of the dendrite using both the motor domain and the C-terminal MT-binding domain.</jats:p>

DOI： 10.7554/elife.00133

その他リンク： https://cdn.elifesciences.org/articles/00133/elife-00133-v1.pdf

生体分子の電子顕微鏡法による 3 次元再構成法

安永卓生, 我妻竜三

顕微鏡 ( 日本顕微鏡学会 ) 47 ( 2 ) 110 - 117 2012年06月

Tilt-angle measurement of a sample stage using a capacitive liquid-based inclinometer 査読有り

Kyoko Matoba, Junichi Takagi, Takuo Yasunaga, Hiroshi Jinnai, Kenji Iwasaki

Microscopy 61 ( 3 ) 193 - 198 2012年06月

記述言語：英語掲載種別：研究論文（学術雑誌）

Alignment of projection images in tomographic reconstruction is a critical process that governs the quality of the reconstructed three-dimensional (3D) image. The most popular alignment method is the marker-based alignment, which typically uses colloidal gold particles added to the specimen (called fiducial markers) to calculate the coordinates of each projection image in the tilt series. This method, however, is not effective when each image contains only a small number of fiducial markers. Therefore, of all the parameters required for alignment, we focussed on the tilt angle and attempted to gage it directly in order to examine whether the acquired angle is accurate enough to perform tomographic reconstruction. We showed that the tilt angle measured using a commercially available capacitive liquid-based inclinometer is more precise than the reading from the monitor of the electron microscope and that it can lead to 3D reconstructions of quality similar to those obtained by the marker-based alignment method. The Author 2012. Published by Oxford University Press [on behalf of Japanese Society of Microscopy]. All rights reserved.

DOI： 10.1093/jmicro/dfs034

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84862898328&origin=inward

Enhanced detection efficiency of genetically encoded tag allows the visualization of monomeric proteins by electron microscopy 査読有り

Yuko Fukunaga, Ai Higashihara, Yuri Nishino, Takuo Yasunaga, Mingyue Jin, Atsuo Miyazawa

Journal of Electron Microscopy 61 ( 4 ) 229 - 236 2012年04月

記述言語：英語掲載種別：研究論文（学術雑誌）

A cadmium-binding, genetically encoded protein tag, consisting of three repeats of metallothionein (3MT), can be used in electron microscopy for the visualization of multimeric- but not monomeric-tagged proteins due to insufficient electron density in monomeric proteins. Here, we present a technique for detecting monomeric 3MT-tagged green fluorescent protein (GFP-3MT) using a platinum compound to intensify the electron density. Substitution of cadmium by platinum as a result of incubating purified cadmium-binding 3MT-tagged GFP (GFP-Cd-3MT) with cis-diamminedichloroplatinum(II) (cisDDP) was assessed by a UV absorption band centered at 284 nm thereby indicating platinum-sulfhydryl bonds. The incubation time and the concentration of cisDDP to reach maximal absorption were 2 h and 36-fold molar equivalent of cisDDP, respectively. GFP-Pt-3MT isolated by gel filtration chromatography contained 29 platinum atoms per single GFP-3MT molecule. Electron-dense particles were observed in a GFP-Pt-3MT sample by electron microscopy without negative staining. Further image processing and image analysis demonstrated that particles with higher density relative to their surroundings were detectable in both GFP-Cd-3MT and GFP-Pt-3MT samples. These results demonstrate that replacement of cadmium with platinum, together with proper image analyses, improve detection efficiency and enable the visualization of 3MT-tagged monomeric protein by electron microscopy. © 2012 The Author. Published by Oxford University Press.

DOI： 10.1093/jmicro/dfs043

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84864976128&origin=inward

1PT185 拘束付き分子動力学法を適用した、電子顕微鏡法のための原子モデル構築ツールの開発(日本生物物理学会第50回年会(2012年度))

Yamashita Risa, Nakao Ryota, Sakamoto Hiroshi, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 52 ( supplement ) S100 2012年01月

StHsp14.0, a small heat shock protein of Sulfolobus tokodaii strain 7, protects denatured proteins from aggregation in the partially dissociated conformation 査読有り

Abe T., Oka T., Nakagome A., Tsukada Y., Yasunaga T., Yohda M.

JOURNAL OF BIOCHEMISTRY 150 ( 4 ) 403 - 409 2011年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

The small heat shock protein (sHsp), categorized into a class of molecular chaperones, binds and stabilizes denatured proteins for the purpose of preventing aggregation. The sHsps undergo transition between different oligomeric states to control their nature. We have been studying the function of sHsp of Sulfolobus tokodaii, StHsp14.0. StHsp14.0 exists as 24meric oligomer, and exhibits oligomer dissociation and molecular chaperone activity over 80°C. We constructed and characterized StHsp14.0 mutants with replacement of the C-terminal IKI to WKW, IKF, FKI and FKF. All mutant complexes dissociated into dimers at 50°C. Among them, StHsp14.0FKF is almost completely dissociated, probably to dimers. All mutants protected citrate synthase (CS) from thermal aggregation at 50°C. But, the activity of StHsp14.0FKF was the lowest. Then, we examined the complexes of StHsp14.0 mutants with denatured CS by SAXS. StHsp14.0WKW protects denatured CS by forming the globular complexes of 24 subunits and a substrate. StHsp14.0FKF also formed similar complex but the number of subunits in the complex is a little smaller. These results suggest that the dimer itself exhibits low chaperone activity, and a partially dissociated oligomer of StHsp14.0 protects a denatured protein from interacting with other molecules by surrounding it. © 2011 The Authors. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

DOI： 10.1093/jb/mvr074

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=80053558864&origin=inward

1G1312 3D structure of the 26S proteasome by high-resolution electron cryo-microscopy(Protein: Structure 1,The 49th Annual Meeting of the Biophysical Society of Japan)

Okuma Yasuo, Kasuya Daisuke, Mitsuoka Kaoru, Saeki Yasushi, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 51 ( 0 ) S45 2011年01月

1G1424 冷陰極電界放出銃を用いた、クライオ電子顕微鏡法による小さい生体分子複合体構造解析への挑戦(蛋白質_構造 1,第49回日本生物物理学会年会)

Watanabe Keita, Ogawa Kaori, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 51 ( 0 ) S46 2011年01月

Structure and Function of Small Heat Shock Proteins from the Magnetotactic Bacterium Magnetospirillum magneticum AMB-1 査読有り

Abe T, Ito S, Nishi N, Tsukada Y, Yasunaga T, Araki A, Matsunaga T, Yohda M,

KOBUNSHI RONBUNSHU ( 公益社団法人高分子学会 ) 67 ( 12 ) 698 - 704 2010年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

スモールヒートショックプロテイン(sHsp)は通常の温度条件下では分子量 12-43 kDa のサブユニットが大きな複合体を形成しているが，熱ストレス下においてはその構造が可逆的に解離する．この解離が sHsp の機能発現に重要な役割をもつと考えられている．磁性細菌 Magnetospirillum magneticum AMB-1 には sHsp とアミノ酸配列の相同性がある ORF が三つ存在した(YP_423206，YP_423303，YP_421578)．分子量からそれぞれ，Hsp17.8, Hsp17.5, Hsp18.3 と命名し大腸菌により大量発現させ精製した．それぞれ，モデル基質である豚心臓由来クエン酸合成酵素の凝集を抑制し，分子シャペロンとして機能していることが明らかとなった．また，ゲルろ過クロマトグラフィーや電子顕微鏡観察により Hsp17.8 はこれまでに報告のある球状のオリゴマーを形成し，Hsp17.5，Hsp18.3 は興味深いことに，巨大なフィラメント構造を形成していた．これまでにも，sHsp がフィラメント構造を形成する報告はあるものの，タンパク質の凝集抑制能を有するものとしては，初めての例である．

DOI： 10.1295/koron.67.698

Structural Basis for Actin Assembly, Activation of ATP Hydrolysis, and Delayed Phosphate Release 査読有り

Murakami K., Yasunaga T., Noguchi TQP, Gomibuchi Y., Ngo KX, Uyeda TQP, Wakabayashi T.

CELL 143 ( 2 ) 275 - 287 2010年10月

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）

Assembled actin filaments support cellular signaling, intracellular trafficking, and cytokinesis. ATP hydrolysis triggered by actin assembly provides the structural cues for filament turnover in vivo. Here, we present the cryo-electron microscopic (cryo-EM) structure of filamentous actin (F-actin) in the presence of phosphate, with the visualization of some α-helical backbones and large side chains. A complete atomic model based on the EM map identified intermolecular interactions mediated by bound magnesium and phosphate ions. Comparison of the F-actin model with G-actin monomer crystal structures reveals a critical role for bending of the conserved proline-rich loop in triggering phosphate release following ATP hydrolysis. Crystal structures of G-actin show that mutations in this loop trap the catalytic site in two intermediate states of the ATPase cycle. The combined structural information allows us to propose a detailed molecular mechanism for the biochemical events, including actin polymerization and ATPase activation, critical for actin filament dynamics. © 2010 Elsevier Inc.

DOI： 10.1016/j.cell.2010.09.034

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=77957793677&origin=inward

Three Outer Arm Dynein Heavy Chains of Chlamydomonas reinhardtii Operate in a Coordinated Fashion Both In Vitro and In Vivo 査読有り

Takazaki H, Liu ZM, Jin MY, Kamiya R, Yasunaga T

CYTOSKELETON 67 ( 7 ) 466 - 476 2010年07月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Outer arm dynein (OAD) in cilia and flagella contains two to three nonidentical heavy chains (HCs) that possess motor activity. In Chlamydomonas, flagellar OAD contains three HCs, a-, b-, and c-HCs, each appearing to have a distinct role. To determine the precise molecular mechanism of their function, cross-sectional electron micrographs of wild-type and single HC-disruption mutants were compared and statistically analyzed. While the a-HC mutant displayed an OAD of lower density, which was attributed to a lack of a-HC, the OAD of band c-HC mutants not only lacked the corresponding HC, but was also significantly affected in its structure, particularly with respect to the localization of a-HC. The lack of b-HC induced mislocalization of a-HC, while a disruption of the c-HC gene resulted in the synchronized movement of a-HC and b-HC in the manners for stacking. Interestingly, using cryo-electron microscopy, purified OADs were typically observed consisting of two stacked heads and an independent single head, which presumably corresponded to c-HC. This conformation is different from previous reports in which the three HCs displayed a stacked form in flagella observed by cryo-electron tomography and a bouquet structure on mica in deep-etch replica images. These results suggest that c-HC supports the tight stacking arrangement of inter or intra a-/b-HC to facilitate the proper functioning of OAD. © 2010 Wiley-Liss, Inc.

DOI： 10.1002/cm.20459

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=77955867545&origin=inward

1P150 1D1435 アクチン重合とそれに伴うATPase活性化の構造的基盤(筋肉(筋蛋白質・収縮),口頭発表,第48回日本生物物理学会年会)

Murakami Kenji, Yasunaga Takuo, Noguchi Taro, Gomibuchi Yuki, Ngo Kien, Uyeda Taro, Wakabayashi Takeyuki

生物物理 ( 一般社団法人日本生物物理学会 ) 50 ( supplement2 ) S45 - S46 2010年01月

1P325 小さい生体分子複合体のためのクライオ電子顕微鏡法の挑戦(バイオイメージング,第48回日本生物物理学会年会)

Watanabe Keita, Ogawa Kaori, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 50 ( supplement2 ) S77 2010年01月

1P166 クラミドモナスの外腕ダイニンが持つ構造特徴(分子モーター,第48回日本生物物理学会年会)

Takazaki Hiroko, Jin Mingyue, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 50 ( supplement2 ) S48 2010年01月

1P168 クライオ電子顕微鏡法による車軸藻ミオシンの高速移動分子メカニズムの解明(分子モーター,第48回日本生物物理学会年会)

Hidaka Hiroaki, Mine Hiroshi, Ito Kohji, Yamamoto Keiichi, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 50 ( supplement2 ) S49 2010年01月

1P153 硬直状態におけるアクチン-ミオシン硬直複合体の三次元構造(筋肉(筋蛋白質・収縮),第48回日本生物物理学会年会)

Tsukada Yoshihiro, Wakabayashi Takeyuki, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 50 ( supplement2 ) S46 2010年01月

Dynein pulls microtubules without rotating its stalk 査読有り国際誌

Ueno H., Yasunaga T., Shingyoji C., Yamaguchi T., Hirose K.

Proceedings of the ASME Summer Bioengineering Conference 2009, SBC2009 ( PART A ) 555 - 556 2009年12月

1P-114 アクチン-ミオシン硬直複合体の高分解能3次元像の獲得(筋肉(筋蛋白質・収縮),第47回日本生物物理学会年会)

Tsukada Yoshihiro, Wakabayashi Takeyuki, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 49 ( supplement ) S81 2009年01月

1P-133 軸糸外腕ダイニンの構造及び微小管と複合体を形成する際のメカニズム(分子モーター,第47回日本生物物理学会年会)

Jin Mingyue, Takazaki Hiroko, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 49 ( supplement ) S84 2009年01月

1P-136 カタユウレイボヤ軸糸外腕ダイニンの構造解析(分子モーター,第47回日本生物物理学会年会)

Harano Jun, Ueno Hironori, Mizuno Katsutoshi, Yasunaga Takuo, Inaba Kazuo, Hirose Keiko

生物物理 ( 一般社団法人日本生物物理学会 ) 49 ( supplement ) S84 2009年01月

Dynein pulls microtubules without rotating its stalk 査読有り国際誌

Ueno H., Yasunaga T., Shingyoji C., Hirose K.

Proceedings of the National Academy of Sciences of the United States of America ( Proceedings of the National Academy of Sciences ) 105 ( 50 ) 19702 - 19707 2008年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Dynein is a microtubule motor that powers motility of cilia and flagella. There is evidence that the relative sliding of the doublet microtubules is due to a conformational change in the motor domain that moves a microtubule bound to the end of an extension known as the stalk. A predominant model for the movement involves a rotation of the head domain, with its stalk, toward the microtubule plus end. However, stalks bound to microtubules have been difficult to observe. Here, we present the clearest views so far of stalks in action, by observing sea urchin, outer arm dynein molecules bound to microtubules, with a new method, "cryopositive stain" electron microscopy. The dynein molecules in the complex were shown to be active in in vitro motility assays. Analysis of the electron micrographs shows that the stalk angles relative to microtubules do not change significantly between the ADP·vanadate and no-nucleotide states, but the heads, together with their stalks, shift with respect to their A-tubule attachments. Our results disagree with models in which the stalk acts as a lever arm to amplify structural changes. The observed movement of the head and stalk relative to the tail indicates a new plausible mechanism, in which dynein uses its stalk as a grappling hook, catching a tubulin subunit 8 nm ahead and pulling on it by retracting a part of the tail (linker). © 2008 by The National Academy of Sciences of the USA.

DOI： 10.1073/pnas.0808194105

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=58149376460&origin=inward

Structural basis for tropomyosin overlap in thin (actin) filaments and the generation of a molecular swivel by troponin-T 査読有り国際誌

Kenji Murakami, Murray Stewart, Kayo Nozawa, Kumiko Tomii, Norio Kudou, Noriyuki Igarashi, Yasuo Shirakihara, Soichi Wakatsuki, Takuo Yasunaga, Takeyuki Wakabayashi

Proceedings of the National Academy of Sciences ( Proceedings of the National Academy of Sciences ) 105 ( 20 ) 7200 - 7205 2008年05月

記述言語：英語掲載種別：研究論文（学術雑誌）

<jats:p>Head-to-tail polymerization of tropomyosin is crucial for its actin binding, function in actin filament assembly, and the regulation of actin-myosin contraction. Here, we describe the 2.1 Å resolution structure of crystals containing overlapping tropomyosin N and C termini (TM-N and TM-C) and the 2.9 Å resolution structure of crystals containing TM-N and TM-C together with a fragment of troponin-T (TnT). At each junction, the N-terminal helices of TM-N were splayed, with only one of them packing against TM-C. In the C-terminal region of TM-C, a crucial water in the coiled-coil core broke the local 2-fold symmetry and helps generate a kink on one helix. In the presence of a TnT fragment, the asymmetry in TM-C facilitates formation of a 4-helix bundle containing two TM-C chains and one chain each of TM-N and TnT. Mutating the residues that generate the asymmetry in TM-C caused a marked decrease in the affinity of troponin for actin-tropomyosin filaments. The highly conserved region of TnT, in which most cardiomyopathy mutations reside, is crucial for interacting with tropomyosin. The structure of the ternary complex also explains why the skeletal- and cardiac-muscle specific C-terminal region is required to bind TnT and why tropomyosin homodimers bind only a single TnT. On actin filaments, the head-to-tail junction can function as a molecular swivel to accommodate irregularities in the coiled-coil path between successive tropomyosins enabling each to interact equivalently with the actin helix.</jats:p>

DOI： 10.1073/pnas.0801950105

その他リンク： https://pnas.org/doi/pdf/10.1073/pnas.0801950105

Dynein pulls microtubules without rotating its stalk 査読有り

Ueno H.,Yasunaga T.,Shigyoji,C.,Hirose K

ProNAS 105 19702 - 19707 2008年04月

Evaluation of a 2k CCD camera with an epitaxially-grown CsI scintillator for recording energy-filtered electron cryo-micrographs., Journal of Electron Microscopy, 57(3), 101-112 (2008) 査読有り

Yasunaga,T.,Wakabayashi,T.

Journal of Electron Microscopy 57 ( 3 ) 101 - 112 2008年04月

Partially functional outer arm dynein in a novel Chlamydomonas mutant expressing a truncated γ heavy chain 査読有り

Liu Z.,Takazaki H.,Nakazawa Y.,Sakato M.,Yagi T.,Yasunaga T.,King S.M.,Kamiya R.

Eukaryotic Cell 7 1136 - 1145 2008年04月

Structural basis for tropomyosin overlap in thin (actin) filaments and the generation of a molecular swivel by troponin-T 査読有り

Murakami K.,Stewart M.,Nozawa K.,Tomii K.,Kudou N.,Igarashi N.,Shirakihara Y.,Wakatsuki S.,Yasunaga T.,Wakabayashi T.

ProNAS 105 ( 20 ) 7200 - 7205 2008年04月

Expression of excess receptors and negative feedback control of signal pathways are required for rapid activation and prompt cessation of signal transduction 査読有り

Kobayashi,H.,Azuma,R.,Yasunaga T.

Cell Communication and Signaling 7 ( 3 ) 2008年03月

ナノイメージングを可能にする顕微鏡法・電子顕微鏡を用いたナノイメージング招待有り

安永卓生

ナノイメージング((株)NTS) 2008年01月

1P-143 ダイニン・微小管複合体の構造学的研究(分子モーター(1),第46回日本生物物理学会年会)

Ueno Hironori, Yasunaga Takuo, Shingyoji Chikako, Hirose Keiko

生物物理 ( 一般社団法人日本生物物理学会 ) 48 ( supplement ) S43 2008年01月

第一編ナノイメージングを可能にする顕微鏡法・第3章電子顕微鏡を用いたナノイメージング招待有り

安永卓生

ナノイメージング((株)NTS) 15 2008年01月

A genetically　encoded metallothionein tag enabling efficient protein detection by electron microscopy 査読有り

Nishino Y,Yasunaga T,Miyazawa A

Journal of Electron Microscopy 56 ( 3 ) 93 - 101 2007年08月

A genetically encoded metallothionein tag enabling efficient protein detection by electron microscopy 査読有り

Nishino Y., Yasunaga T., Miyazawa A.

Journal of Electron Microscopy 56 ( 3 ) 93 - 101 2007年06月

記述言語：英語掲載種別：研究論文（学術雑誌）

The observation of biological materials by transmission electron microscopy (TEM) frequently requires the use of negative staining and/or immuno-gold labeling to visualize or identify the specimen, but these techniques are limited. In contrast, genetic labeling with green fluorescent protein (GFP) and its homologs have led to rapid advances in the observation of proteins of interest in cells by light microscopy (LM). These fluorescent tags allow for the visualization of dynamic processes in live cells without the use of secondary reagents. Here, we report the development of an artificial metalloprotein fusion protein expressed in Escherichia coli grown in Cd2+-containing medium that allows for efficient protein detection by TEM without additional staining steps. We linked the subunits of the bacterial 14-mer protein GroEL with three repeats of metallothionein (3MT). The 3MT-fused GroEL (GroEL-14(3MT)) was successfully expressed in E. coli, and the purified protein included 250 cadmium atoms per molecule on average. Cd2+-bound GroEL-14(3MT) was detected by TEM in the absence of negative staining on a carbon grid, and the particle densities of GroEL-14(3MT) were much greater than those of untagged GroEL in vitreous ice. Taken together, our data indicate that the 3MT tag provides a promising means of allowing the identification of oligomeric proteins isolated from cells in the absence of other detection techniques. © The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved.

DOI： 10.1093/jmicro/dfm008

その他リンク： https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=35948943072&origin=inward

Domains 16 and 17 of tropoelastin in elastic fiber formation 査読有り

Wachi H.,Sato F.,Nakazawa J.,Nonaka R.,Szabo Z.,Urban Z.,Yasunaga T.,Maeda I.,Okamoto K.,Starcher B.C.,Li D.Y.,Mecham R.P.,Seyama Y.

Biochemical Journal. 402 60 - 70 2007年04月

An actin homolog of the archaeon Thermoplasma acidophilum that retains the ancient trait of eukaryotic actin 査読有り

Hara F.,Yamashiro K.,Nemoto N.,Ohta Y,Yokobori S,Yasunaga T.,Hisanaga S. and Yamagishi A

J. Bacteriology. 189 ( 5 ) 2039 - 2045 2007年04月

電子顕微鏡画像解析ソフトウェア統合環境Eosの試み招待有り査読有り

安永　卓生

顕微鏡 42 ( 3 ) 2007年04月

1P054 単粒子解析のための粒子投影方向決定の新手法 : シノグラムライン特徴量とその応用(蛋白質(計測解析の方法論),ポスター発表,第45回日本生物物理学会年会)

村山涼子, 安永卓生

生物物理 ( 一般社団法人日本生物物理学会 ) 47 ( supplement ) S37 2007年01月

電子顕微鏡画像解析ソフトウェア統合環境査読有り

安永卓生

Eosの試み 42 214 - 216 2007年01月

1P055 多種の処理ツールの統合による半自動3次元再構成システム(蛋白質(計測解析の方法論),ポスター発表,第45回日本生物物理学会年会)

新名人士, 谷口佳苗, 西野有里, 岩崎憲治, 高本淳一, 宮澤敦夫, 安永卓生

生物物理 ( 一般社団法人日本生物物理学会 ) 47 ( supplement ) S37 2007年01月

電子顕微鏡法の新展開招待有り

安永　卓生

現代化学 ( 7 ) 2006年07月

Atomic model construction of protein complexes from electron micrographs and visualization of their 3D structure using VR system 査読有り

Noda K,Nakamura M,Nishida R,Yoneda Y,Yamaguchi Y,TAMURA Y,Nakuamura H.,Yasunaga T

J. Plasma Physics 72 ( 06 ) 1037 - 1040 2006年06月

Comparative analysis of the protein folding activities of two chaperonin subunits of Thermococcus strain KS-1; the effects of beryllium fluoride 査読有り

Yoshida T.,Iizuka R.,Itami K.,Yasunaga T.,Sakuraba H.,Ohshima T.,Yohda M.,Maruyama T.

Extremophiles 11 ( 2 ) 225 - 235 2006年04月

19 Thermoplasma acidophilum由来アクチンの特性解明および真核生物アクチンとの比較検討(一般講演,第31回学術講演会講演要旨集)

原太志, 山城寛, 根本直樹, 太田好則, 安永卓生, 久永真市, 山岸明彦

Viva origino ( 文部省科学研究費総合研究(B)「原始地球上における生命の起源に関する研究」班 ) 34 ( 1 ) 20 2006年03月

電子顕微鏡法の新展開招待有り

安永卓生

現代化学 424号 2006年01月

1P230 Visualization of the ATP Hydrolysis Intermediates of Myosin II by Electron Cryo-microscopy(8. Muscle contraction and muscle protein,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Ohta Yoshinori, Umeki Nobuhisa, Katayama Eisaku, Maruta Shinsaku, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S204 2006年01月

1P575 Potential structure changes of dynein stalk by molecular dynamics calculation(27. Molecular dynamics simulation,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Fukae Kazuki, Sutoh Kazuo, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S290 2006年01月

1P523 Dvelopment of novel automated determination of contrast transfer function of electron microscopy(25. New methods and tools (I),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Imamura Motoki, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S277 2006年01月

1P522 Fabrication of freezing apparatus with controlling temperature and humidity for electron cryo-microscopy(25. New methods and tools (I),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Hirai Chie, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S277 2006年01月

1P286 Analysis of unconventional myosins by spectroscopic electron cryomicroscopy.(9. Molecular motor (I),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Fujiwara Shingo, Kano Yousuke, Sato Osamu, Honma Kazuaki, Ikebe Reiko, Ikebe Mitsuo, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S218 2006年01月

1P267 Structural study of sea urchin outer-arm dynein bound to microtubules(9. Molecular motor (I),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Ueno Hironori, Yoshimura Azumi, Yasunaga Takuo, Shingyoji Chikako, Hirose Keiko

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S213 2006年01月

1P265 Localization of Three Chlamydomonas Outer Arm Heavy Chains(9. Molecular motor (I),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

Takazaki Hiroko, Liu Zhongmei, Kamiya Ritsu, Yasunaga Takuo

生物物理 ( 一般社団法人日本生物物理学会 ) 46 ( supplement2 ) S213 2006年01月

Atomic model construction of protein complexes from electron micrographs and visualization of their 3D structure using VR system 査読有り

Yasunaga T.,Noda K.,Nakamura M.,Nishida R.,Yamaguchi Y.,Tamura Y.,Nakamura,H.

Joint conference of 19th International. Conference on Numerical Simulation. of Plasmas (ICNSP) and 7th Asia. Pacific Plasma Theory Conference 2005年07月

An engineered chaperonin cagin a guest protein: Structural insights and potential as a protein expression tool 査読有り

Furutani M.,Hata J.,Shomura Y.,Itami K.,Yoshida T.,Izumoto Y.,Togi A.,Ideno A.,Yasunaga T.,Miki K.,Murayama T.

Protein Science 14 ( 2 ) 341 - 350 2005年04月

An axonemal dynein particularly important for flagellar movement at high viscosity: Implications from a new chlamydomonas mutant deficient in the dynein heavy chain gene DHC9 査読有り

Yagi T.,Minoura I,Fujiwara A.,Saito R.,Yasunaga T.,Hirono M.,Kamiya R.

J. Biol. Chem. 280 41412 - 41420 2005年04月

Structural basis for Ca2+-regulated muscle relaxation at interaction sites of troponin with actin and tropomyosin 査読有り

Murakami K,Yumoto F,Ohki SY,Yasunaga T,Tanokura M,Wakabayashi T.

J Mol Biol. 352 ( 1 ) 178 - 201 2005年04月

Automatic Detecting Particles Approach Using Adaboost-SVM 査読有り

Yang J.,Ohashi T.,& Yasunaga T.

Wseas Transactinns on Biology and Biomedicine 2 163 - 168 2005年04月

Automatic Detecting particles Approach Using Adaboost-SVM 査読有り

Yang J.,Ohashi T.,Yasunaga T.

WSEAS Conferences (MCBC(Mathematics and Computers in Biology and Chemistry)’05) 2005年03月

Automatic Actomyosin complex selection using SVM 査読有り

Yang J.,Ohashi T.,Yasunaga T.

IADIS International Conference: Applied Computing 2005 2005年02月

A visualization tool of biological macromolecular structures toward in slico microscopy

安永卓生

Annual Report of NIFS 2002 2003年03月

Archaeal Group II Chaperonin Mediates Protein Folding in the cis-Cavity Without a Detachable GroES-like Co-chaperonin 査読有り

Yoshida T.,Kawaguchi R.,Taguchi H.,Yoshida M.,Yasunaga T.,Wakabayashi T.,Yohda M.,Maruyama T

J. Mol. Biol. 315 ( 1 ) 73 - 85 2002年01月

Archaeal group II chaperonin mediates protein folding in the cis-cavity without a detachable GroES-like co-chaperonin 査読有り国際誌

Yoshida T., Kawaguchi R., Taguchi H., Yoshida M., Yasunaga T., Wakabayashi T., Yohda M., Maruyama T.

Journal of Molecular Biology 315 ( 1 ) 73 - 85 2002年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

Group II chaperonins of archaea and eukaryotes are distinct from group I chaperonins of bacteria. Whereas group I chaperonins require the co-chaperonin Cpn-10 or GroES for protein folding, no co-chaperonin has been known for group II. The protein folding mechanism of group II chaperonins is not yet clear. To understand this mechanism, we examined protein refolding by the recombinant α or β-subunit chaperonin homo-oligomer (α16mer and β16mer) from a hyperthermoplilic archaeum, Thermococcus strain KS-1, using a model substrate, green fluorescent protein (GFP). The α16mer and β16mer captured the non-native GFP and promoted its refolding without any co-chaperonin in an ATP dependent manner. A non-hydrolyzable ATP analog, AMP-PNP, induced the GFP refolding mediated by β16mer but not by the α16mer. A mutant α-subunit chaperonin homo-oligomer (trap-α) could capture the non-native protein but lacked the ability to refold it. Although trap-α suppressed ATP-dependent refolding of GFP mediated by α16mer or β16mer, it did not affect the AMP-PNP-dependent refolding. This indicated that the GFP refolding mediated by β16mer with AMP-PNP was not accessible to the trap-α. Gel filtration chromatography and a protease protection experiment revealed that this refolded GFP, in the presence of AMP-PNP, was associated with β16mer. After the completion of GFP refolding mediated by β16mer with AMP-PNP, addition of ATP induced an additional refolding of GFP. Furthermore, the β16mer preincubated with AMP-PNP showed the ability to capture the non-native GFP. These suggest that AMP-PNP induced one of two chaperonin rings (cis-ring) to close and induced protein refolding in this ring, and that the other ring (trans-ring) could capture the unfolded GFP which was refolded by adding ATP. The present data indicate that, in the group II chaperonin of Thermococcus strain KS-1, the protein folding proceeds in its cis-ring in an ATP-dependent fashion without any co-chaperonin. © 2002 Academic Press.

DOI： 10.1006/jmbi.2001.5220