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Structural fluctuations of protein complexes measured by combination of protein engineering and solvent quenching meshod

Research Project

Project/Area Number 23770194
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Biophysics
Research InstitutionYamagata University (2012-2013)
National Institutes of Natural Sciences Okazaki Research Facilities (2011)

Principal Investigator

MAKABE koki  山形大学, 理工学研究科, 准教授 (20508072)

Project Period (FY) 2011 – 2013
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywordsタンパク質工学 / シャペロン / 水素交換 / GroEL / 蛋白質 / 構造揺らぎ / タンパク質 / 水素重水素交換
Research Abstract

In this project, we have been studying the structural fluctuations of E. coli chaperonin GroEL/ES system by using DMSO-quenched H/D exchange methods. For GroES H/D exchange experiments, we have successfully determined the exchange rates for 33 amide protons out of 94 amide protons in GroES. For the remaining 61 amide protons, we have determined the lower limit of the exchange rates. We found that only 10 amide protons have a protection factor larger than 10 to the power 6th. This result indicates that the free GroES is highly flexible in significant portions. From crystal structure analysis of N-terminal mutants of alpha-lactalbumin, a substrate protein of GroEL/ES system, we found the hydrogen bonding of the N-terminal residue is important for its stability.

Report

(4 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • 2011 Research-status Report
  • Research Products

    (32 results)

All 2014 2013 2012 2011 Other

All Journal Article (23 results) (of which Peer Reviewed: 23 results) Presentation (5 results) (of which Invited: 3 results) Remarks (1 results) Patent(Industrial Property Rights) (3 results)

  • [Journal Article] Dissection of the ATP-dependent conformational change cycle of a group II chaperonin.2014

    • Author(s)
      Nakagawa A, Moriya K, Arita M, Yamamoto Y, Kitamura K, Ishiguro N, Kanzaki T, Oka T, Makabe K, Kuwajima K, Yohda M.
    • Journal Title

      J Mol Biol.

      Volume: 426 Issue: 2 Pages: 447-459

    • DOI

      10.1016/j.jmb.2013.09.034

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The catalytic domain of topological knot tRNA methyltransferase (TrmH) discriminates between substrate tRNA and non-substrate tRNA via an induced-fit process2013

    • Author(s)
      Ochi A, Makabe K, Yamagami R, Hirata A, Sakaguchi R, Hou YM, Watanabe K, Nureki O, Kuwajima K, Hori H
    • Journal Title

      J Biol Chem

      Volume: 30

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] ATP dependent rotational motion of group II chaperonin observed by X-ray single molecule tracking2013

    • Author(s)
      Sekiguchi H, Nakagawa A, Moriya K, Makabe K, Ichiyanagi K, Nozawa S, Sato T, Adachi S, Kuwajima K, Yohda M, Sasaki YC
    • Journal Title

      PLoS One

      Volume: 29

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] NMR characterization of the interaction of GroEL with amyloidβas a model ligand2013

    • Author(s)
      Yagi-Utsumi M, Kunihara T, Nakamura T, Uekusa Y, Makabe K, Kuwajima K, Kato K
    • Journal Title

      FEBS Lett

      Volume: 587 Pages: 1605-9

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] The H/D-exchange Kinetics of the Escherichia coli Co-chaperonin GroES Studied by 2D-NMR and DMSO-Quenched Exchange Methods2013

    • Author(s)
      Chandak MS, Nakamura T, Makabe K, Takenaka T, Mukaiyama A, Chaudhuri TK, Kato K, Kuwajima K
    • Journal Title

      J Mol Biol

      Volume: 425 Pages: 2541-60

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Molecular mechanisms of the cytotoxicity of humanα-lactalbumin made lethal to tumor cells (HAMLET) and other protein-oleic acid complexes2013

    • Author(s)
      Nakamura T, Aizawa T, Kariya R, Okada S, Demura M, Kawano K, Makabe K, Kuwajima K
    • Journal Title

      J Biol Chem

      Volume: 288 Pages: 14408-14416

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural insights into the stability perturbations induced by N-terminal variation in human and goatα-lactalbumin2013

    • Author(s)
      Makabe K, Nakamura T, Kuwajima K
    • Journal Title

      Protein Eng Des Sel

      Volume: 26 Pages: 165-170

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] The Molten Globule ofβ2-Microglobulin Accumulated at pH 4 and Its Role in Protein Folding2013

    • Author(s)
      Mukaiyama A, Nakamura T, Makabe K, Maki K, Goto Y, Kuwajima K
    • Journal Title

      J Mol Biol

      Volume: 425 Pages: 273-291

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Native-state Heterogeneity ofβ2-Microglobulin as Revealed by Kinetic Folding and Real-time NMR Experiments2013

    • Author(s)
      Mukaiyama A, Nakamura T, Makabe K, Maki K, Goto Y, Kuwajima K
    • Journal Title

      J Mol Biol

      Volume: 425 Pages: 257-272

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural insights into the stability perturbations induced by N-terminal variation in human and goat α-lactalbumin.2013

    • Author(s)
      Makabe K, Nakamura T, Kuwajima K
    • Journal Title

      Protein Eng Des Sel.

      Volume: 26 Issue: 2 Pages: 165-170

    • DOI

      10.1093/protein/gzs093

    • Related Report
      2013 Annual Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Molecular mechanisms of the cytotoxicity of human α-lactalbumin made lethal to tumor cells (HAMLET) and other protein-oleic acid complexes.2013

    • Author(s)
      Nakamura T, Aizawa T, Kariya R, Okada S, Demura M, Kawano K, Makabe K, Kuwajima K.
    • Journal Title

      J Biol Chem.

      Volume: 288 Issue: 20 Pages: 14408-14416

    • DOI

      10.1074/jbc.m112.437889

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The H/D-exchange Kinetics of the Escherichia coli Co-chaperonin GroES Studied by 2D-NMR and DMSO-Quenched Exchange Methods.2013

    • Author(s)
      Chandak MS, Nakamura T, Makabe K, Takenaka T, Mukaiyama A, Chaudhuri TK, Kato K, Kuwajima K.
    • Journal Title

      J Mol Biol.

      Volume: 425 Issue: 14 Pages: 2541-2560

    • DOI

      10.1016/j.jmb.2013.04.008

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] NMR characterization of interaction of GroEL with amyloid β as a model ligand.2013

    • Author(s)
      M.Yagi-Utsumi, T.Kunihara, T.Nakamura, Y.Uekusa, K.Makabe, K.Kuwajima, and K.Kato
    • Journal Title

      FEBS Lett.

      Volume: 587 Issue: 11 Pages: 605-609

    • DOI

      10.1016/j.febslet.2013.04.007

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] ATP dependent rotational motion of group II chaperonin observed by X-ray single molecule tracking.2013

    • Author(s)
      Sekiguchi H, Nakagawa A, Moriya K, Makabe K, Ichiyanagi K, Nozawa S, Sato T, Adachi S, Kuwajima K, Yohda M, Sasaki YC.
    • Journal Title

      PLoS One

      Volume: 8 Issue: 5 Pages: e64176-e64176

    • DOI

      10.1371/journal.pone.0064176

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The catalytic domain of topological knot tRNA methyltransferase (TrmH) discriminates between substrate tRNA and nonsubstrate tRNA via an induced-fit process.2013

    • Author(s)
      A. Ochi, K. Makabe, R. Yamagami, A. Hirata, R. Sakaguchi, Y.M. Hou, K. Watanabe, O. Nureki, K. Kuwajima, and H. Hori
    • Journal Title

      J. Biol. Chem.

      Volume: 288 Issue: 35 Pages: 25562-25574

    • DOI

      10.1074/jbc.m113.485128

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Native-state heterogeneity of β(2)-microglobulin as revealed by kinetic folding and real-time NMR experiments.2013

    • Author(s)
      Mukaiyama A, Nakamura T, Makabe K, Maki K, Goto Y, Kuwajima K.
    • Journal Title

      J Mol Biol.

      Volume: 425 Issue: 2 Pages: 257-272

    • DOI

      10.1016/j.jmb.2012.11.004

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] The molten globule of β(2)-microglobulin accumulated at pH 4 and its role in protein folding2013

    • Author(s)
      A. Mukaiyama, T. Nakamura, K. Makabe, K. Maki, Y. Goto, K. Kuwajima
    • Journal Title

      J. Mol. Biol.

      Volume: 425 Issue: 2 Pages: 273-291

    • DOI

      10.1016/j.jmb.2012.11.002

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Sequential Four-state Folding/unfolding of Goatα-Lactalbumin and Its N-Terminal Variants Proteins2012

    • Author(s)
      Tomoyori K, Nakamura T, Makabe K, Maki K, Saeki K, Kuwajima K
    • Volume
      80
    • Pages
      2191-206
    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Fibrillogenic propensity of the GroEL apical domain : A Janus-faced minichaperone2012

    • Author(s)
      Chen J, Yagi H, Sormanni P, Vendruscolo M, Makabe K, Nakamura T, Goto Y, Kuwajima K
    • Journal Title

      FEBS Letters

      Volume: 586 Pages: 1120-1127

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Sequential four-state folding/unfolding of goat α-lactalbumin and its N-terminal variants.2012

    • Author(s)
      Tomoyori K, Nakamura T, Makabe K, Maki K, Saeki K, Kuwajima K.
    • Journal Title

      Proteins.

      Volume: 80 Issue: 9 Pages: 2191-2206

    • DOI

      10.1002/prot.24109

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Fibrillogenic propensity of the GroEL apical domain: A Janus-faced minichaperone2012

    • Author(s)
      Chen J, Yagi H, Sormanni P, Vendruscolo M, Makabe K, Nakamura T, Goto Y, Kuwajima K.
    • Journal Title

      FEBS Letters

      Volume: 586 Pages: 1120-1127

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] Dissecting a Bimolecular Process of MgATP2- Binding to the Chaperonin GroEL2011

    • Author(s)
      Chen J, Makabe K, Nakamura T, Inobe T, Kuwajima K
    • Journal Title

      J Mol Biol

      Volume: 410 Pages: 343-356

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Dissecting a bimolecular process of MgATP2- binding to the chaperonin GroEL.2011

    • Author(s)
      Chen J, Makabe K, Nakamura T, Inobe T, Kuwajima K.
    • Journal Title

      J Mol Biol

      Volume: 412 Pages: 343-356

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Presentation] Folding and redesign of a beta-sheet rich model protein2013

    • Author(s)
      Koki Makabe
    • Organizer
      International symposium on protein folding and its biological significance
    • Place of Presentation
      岡崎
    • Year and Date
      2013-03-05
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] Folding and redesign of a beta-sheet rich model protein2013

    • Author(s)
      Koki Makabe
    • Organizer
      International symposium on protein folding and its biological significance
    • Place of Presentation
      Okazaki, Japan
    • Related Report
      2012 Research-status Report
    • Invited
  • [Presentation] モデル蛋白質によるβシート構造の再設計2012

    • Author(s)
      真壁幸樹
    • Organizer
      平成24年生物物理学会東北支部会
    • Place of Presentation
      仙台
    • Year and Date
      2012-12-21
    • Related Report
      2013 Final Research Report
  • [Presentation] STRUCTURAL INSIGHTS INTO THE STABILITY PERTURBATIONS BY N-TERMINAL VARIATIONS IN HUMAN AND GOAT ALPHA-LACTALBUMIN2012

    • Author(s)
      Koki Makabe, Takashi Nakamura and Kunihiro Kuwajima
    • Organizer
      Korea-Japan Seminars on Biomolecular Sciences Experiments and Simulations
    • Place of Presentation
      奈良
    • Year and Date
      2012-01-09
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] STRUCTURAL INSIGHTS INTO THE STABILITY PERTURBATIONS BY N-TERMINAL VARIATIONS IN HUMAN AND GOAT ALPHA-LACTALBUMIN2012

    • Author(s)
      Koki Makabe, Takashi Nakamura and Kunihiro Kuwajima
    • Organizer
      Korea-Japan Seminars on Biomolecular Sciences Experiments and Simulations(招待講演)
    • Place of Presentation
      東大寺文化センター(奈良県)
    • Related Report
      2011 Research-status Report
  • [Remarks]

    • URL

      https://makabe.yz.yamagata-u.ac.jp

    • Related Report
      2013 Final Research Report
  • [Patent(Industrial Property Rights)] 蛋白質―化学療法剤複合体及びその製造方法,並びに医薬への応用2012

    • Inventor(s)
      桑島邦博,中村敬,真壁幸樹,岡田誠治
    • Industrial Property Rights Holder
      分子科学研究所、熊本大学
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2012-147492
    • Filing Date
      2012
    • Related Report
      2013 Final Research Report
  • [Patent(Industrial Property Rights)] 球状蛋白質の準安定状態を用いた抗癌細胞作用のある分子の作成2011

    • Inventor(s)
      桑島邦博,中村敬,真壁幸樹
    • Industrial Property Rights Holder
      分子科学研究所
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2011-238611
    • Filing Date
      2011
    • Related Report
      2013 Final Research Report
  • [Patent(Industrial Property Rights)] 蛋白質-脂肪酸複合体及び腫瘍細胞の細胞死誘導薬2011

    • Inventor(s)
      桑島邦博、中村敬、真壁幸樹
    • Industrial Property Rights Holder
      自然科学研究機構
    • Industrial Property Number
      2011-238611
    • Filing Date
      2011
    • Related Report
      2011 Research-status Report

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Published: 2011-08-05   Modified: 2019-07-29  

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