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膜結合型AAA-ATPase p47/p97複合体による細胞膜認識とその制御

Research Project

Project/Area Number 15083206
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionYokohama City University

Principal Investigator

JEREMYR.H. Tame (TAME J.R.H. / J・R・H Tame / JEREMY R・H Tame / JEREMY R.H.Tame)  Yokohama City University, 国際総合科学研究科, 教授 (00336588)

Co-Investigator(Kenkyū-buntansha) 朴 三用  横浜市立大学, 国際総合科学研究科, 准教授 (20291932)
雲財 悟  横浜市立大学, 国際総合科学研究科, 助教 (60336592)
Project Period (FY) 2003 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥80,600,000 (Direct Cost: ¥80,600,000)
Fiscal Year 2008: ¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2007: ¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2006: ¥14,400,000 (Direct Cost: ¥14,400,000)
Fiscal Year 2005: ¥14,400,000 (Direct Cost: ¥14,400,000)
Fiscal Year 2004: ¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2003: ¥7,400,000 (Direct Cost: ¥7,400,000)
KeywordsX線結晶構造解析 / タンパク質 / 膜輸送 / 膜蛋白質 / 蛋白質間相互作用 / 生物物理 / 蛋白質
Research Abstract

(1)TRAPチューブ
E50CのみTRAPに変異導入したものとV69Cのみに変異導入したものをチューブの最小単位となるように作製した。これはTRAP同士がどのように結合しチューブとなるかをX線結晶構造解析により解明するためである。D8H、F32D、E50C、R66H、V69C(以下HCCDH)と、D8H、F32A、E50C、R66H、V69C(以下HCCAH)というように5箇所に変異を導入し、E50とV69に導入したシステインによりS-S結合を形成することを予想し設計した。
(2)TRAP-AT複合体
Bacillus subtilisの細胞質の中では、TRAP活性はanti-TRAPというタンパク質に制御されており、トリプトファンの不足している状態で、TRAPとantiTRAPは複合体を作る。したがって、RNA結合が阻害される。今まではどのようにTRAPとantiTRAPが相互作用しているか明らかではなかった。そのため、我々はこの複合体の結晶化を行い、結晶構造を決定した。
TRAP-antiTRAP複合体のX線結晶構造を決定した。結晶化とともに、TRAPは11量体から12量体になった。結晶対称軸とTRAPリング軸を合わせて、非対称単位が数個の分子しか含まない。それから、通常では大きい複合体は十分に分解能回折データを測ることができないが、今回は高い分解能のデータを測ることができた。

Report

(6 results)
  • 2008 Annual Research Report
  • 2007 Annual Research Report
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (40 results)

All 2009 2008 2007 2006 2005 2004 Other

All Journal Article (29 results) (of which Peer Reviewed: 9 results) Presentation (4 results) Book (1 results) Publications (6 results)

  • [Journal Article] The nature of the TRAP-Anti-TRAP complex2009

    • Author(s)
      Watanabe M, et.al.
    • Journal Title

      Proc Natl Acad Sci USA 106(7)

      Pages: 2176-81

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The structural basis for an essential subunit interaction in influenza virus RNA polymerase2008

    • Author(s)
      Obayashi E, et.al.
    • Journal Title

      Nature 454(7208)

      Pages: 1127-31

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Intersubunit linker length as a modifier of protein stability : crvstal structures and thermostability of mutant TRAP2008

    • Author(s)
      Watanabe M, et.al.
    • Journal Title

      ProteinSci 17(3)

      Pages: 518-26

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Intersubunit linker length as a modifier of protein stability:crystal structures and thermostability of mutant TRAP2008

    • Author(s)
      Watanabe M, Mishima Y, Yamashita I, Park SY, Tame JR, Heddle JG
    • Journal Title

      Protin Sci. 17(3)

      Pages: 518-26

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] EFFect of N-terminal reidues on the structural stability of recmbinant horse L-chain apoferritin in an acidic environment2007

    • Author(s)
      Yoshizwa K, Mishima Y, Park SY, Heddle JG, Tame JR, Iwahori K, Kobayashi M, Yamashita I
    • Journal Title

      J Biochem 142(6)

      Pages: 707-13

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Using the ringshaped protein TRAP to capture and confine gold nanod ots on a surface2007

    • Author(s)
      Heddle JG, Fujiwara I, Yamadaki H, Yoshii S, Nishio K, Addy C, Yamsdhita I, Tame JR
    • Journal Title

      Small 3(11)

      Pages: 1950-6

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallication and preliminary crystallographic studies of the metallogl ycoprotein esterase A4 using a baculovirus expressio system2007

    • Author(s)
      Hiraki T, Shibayama N, Yoon YH, Yun KM, Hamamoto T, Tame JR, Park SY
    • Journal Title

      Acta Crystallogr sect F struct Biol Cryst Commun 63(Pt9)

      Pages: 734-6

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structure and mechanism of HpcG,a hydratase in the homoprotocatech uate degradation pathway of Escherichia coli2007

    • Author(s)
      Izumi A, Rea D, Adachi T, Unzai S, Park SY, Roper DI, Tame JR
    • Journal Title

      J Mol Biol 370(5)

      Pages: 899-911

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Dynamic allostery in the ring protein TRAP2007

    • Author(s)
      Heddle JG, Okajima T, Scott DJ, Akashi S, Psrk SY, Tame JR
    • Journal Title

      J Mol Biol 371(1)

      Pages: 154-67

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Limited tolerance towards folded elements during secretion of the autotransporter Hbp.2007

    • Author(s)
      Jong WS, ten Hagen-Jongman CM, den Blaauwen T, Jan Slotboom D, Tame JR, Wickstrom D, de Gier JW, Otto BR, Luinnk J.
    • Journal Title

      Mol Microbiol. 63(5)

      Pages: 1524-1536

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Nickel binding to NikA : an additional binding site reconciles spectroscopy, calorimetry and crystallography.2007

    • Author(s)
      Addy C, Chara M, Kawai F, Kidera A, Ikeguchi M, Fuchigami S, Osawa M, Shimada I, Park SY, Tame JR, Heddle JG
    • Journal Title

      Acta Crystallogr D Biol Crystallogr. 63(Pt 2)

      Pages: 221-229

    • Related Report
      2006 Annual Research Report
  • [Journal Article] 1.25 A resolution crystal structures of human haemoglobin in the oxy, deoxy and carbonmonoxy forms.2006

    • Author(s)
      Park SY, Yokoyama T, Shibayama N, Shiro Y, Tame JR
    • Journal Title

      J Mol Biol. 360(3)

      Pages: 690-701

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Rounding up : Engineering 12-membered rings from the cyclic 11-mer TRAP.2006

    • Author(s)
      Heddle JG, Yokoyama T, Yamashita I, Park SY, Tame JR
    • Journal Title

      Structure. 14(5)

      Pages: 925-933

    • Related Report
      2006 Annual Research Report
  • [Journal Article] The Swi5-Sfrl complex stimulates Rhp51/Rad51- and Dmcl-mediated DNA strand exchange in vitro.2006

    • Author(s)
      Haruta N, Kurokawa Y, Murayama Y, Akamatsu Y, Unzai S, Tsutsui Y, Iwasaki H
    • Journal Title

      Nat Struct Mol Biol 13(9)

      Pages: 823-830

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Structural basis for protein recognition by B30.2/SPRY domains2006

    • Author(s)
      J-S Woo, H-Y Suh, S-Y Park, B-H Oh
    • Journal Title

      Mol. Cell 24

      Pages: 967-976

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Crystal Structure of penicillin Binding Protein4(dacB)from Escherichia coli, both in the Native Form and Covalently Linked to Various Antibiotics.2006

    • Author(s)
      Kishida, H., Unzai, S., Roper, D.I., Lloyd, A., Park, S.Y., Tame, J.R.
    • Journal Title

      Biochemistry 45(3)

      Pages: 783-792

    • Related Report
      2005 Annual Research Report
  • [Journal Article] R-state Haemoglobin with Low Oxygen Affinity : Crystal Structures of Deoxy Human and Carbonmonoxy Horse Haemoglobin Bound to the Effector Molecule L35.2006

    • Author(s)
      Yokoyama, T., Neya, S., Tsuneshige, A., Yonetani, T., Park, S.Y., Tame, J.R.
    • Journal Title

      Journal of Molecular Biology 356(3)

      Pages: 790-801

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Scoring functions--the first 100 years.2005

    • Author(s)
      Tame, J.R
    • Journal Title

      J.Comput.Aided.Mol.Des 19(6)

      Pages: 445-451

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Characterization of an iron-regulated alpha-enolase of Bacteroides fragilis.2005

    • Author(s)
      Sijbrandi, R., Den Blaauwen, T., Tame, J.R., Oudega, B., Luirink, J., Otto, B.R.
    • Journal Title

      Microbes.Infect. 7(1)

      Pages: 9-18

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Backbone 1H,13C, and 15N assignments of a 56 kDa E.coli nickel binding protein NikA.2005

    • Author(s)
      Rajesh, S., Heddle, J.G, Kurashima-Ito, K., Nietlispach, D., Shirakawa, M., Tame, J.R., Ito, Y.
    • Journal Title

      J.Biomol.NMR 32(2)

      Pages: 177-177

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Crystal structure of hemoglobin protease, a heme binding autotransporter protein from pathogenic Escherichia coli.2005

    • Author(s)
      Otto, B.R., Sijbrandi, R., Luirink, J., Oudega, B., Heddle, J.G., Mizutani, K., Park, S.Y., Tame, J.R.
    • Journal Title

      J.Biol.Chem. 280(17)

      Pages: 17339-17345

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Crystal structure of heme binding protein, an autotransporter hemoglobin protease from pathogenic escherichia coli.2005

    • Author(s)
      Otto, B.R., et al.
    • Journal Title

      Journal of Biological Chemistry (in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Crystal structure of a novel polyisoprenoid-binding protein from Thermus thermophilus HB8.2005

    • Author(s)
      Handa, N., et al.
    • Journal Title

      Protein Science 14(4)

      Pages: 1004-1010

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Characterization of an iron-regulated alpha-enolase of Bacteroides fragilis.2004

    • Author(s)
      Sijbrandi, R., et al.
    • Journal Title

      Microbes and infection 7(1)

      Pages: 9-18

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Crystal structure of a conserved hypothetical protein TT1751 from Thermus thermophilus HB8.2004

    • Author(s)
      Kishishita, S., et al.
    • Journal Title

      Proteins 57(4)

      Pages: 883-887

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Crystal structures of unliganded and half-liganded human hemoglobin derivatives cross-linked between Lys 82betal and Lys 82beta2.2004

    • Author(s)
      Park, S.Y., et al.
    • Journal Title

      Biochemistry 43(27)

      Pages: 87111-87117

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Crystal structure of elongation factor P from Thermus thermophilus HB8.2004

    • Author(s)
      Hanawa-Suetsugu K., et al.
    • Journal Title

      Proc.Natl.Acad.Sci.USA. 101(26)

      Pages: 9595-9600

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Novel mechanisms of pH sensitivity in tuna hemoglobin : a structural explanation of the root effect.2004

    • Author(s)
      Yokoyama, T., et al.
    • Journal Title

      Journal of Biological Chemistry 279(27)

      Pages: 28632-28640

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Crystal structures of catalytic domains of pseudouridine synthases RluC and RluD from Escherichia coli.2004

    • Author(s)
      Mizutani, K., et al.
    • Journal Title

      Biochemistry 43(15)

      Pages: 4454-4463

    • Related Report
      2004 Annual Research Report
  • [Presentation] What a crystal structure doesn't tell you2008

    • Author(s)
      Jeremy R.H. Tame
    • Organizer
      よこはまNMR構造生物学研究会国際シンポジウム
    • Place of Presentation
      横浜
    • Related Report
      2008 Annual Research Report
  • [Presentation] Structural insights into serine protease autotoransporter HBP rfom path ogentic Escherichia coli2008

    • Author(s)
      J.Tame
    • Organizer
      Society for General Microbiology
    • Place of Presentation
      Edinburgh,UK
    • Related Report
      2007 Annual Research Report
  • [Presentation] X-ray Crystallography of Autotransporters from Pathogenic Bacteria2007

    • Author(s)
      Kaoru Nishimura, Sam-Yog Park, and Jeremy Tame
    • Organizer
      Molecular Microbiology Young Microbiologists Minisymposium Prokaryotic Membranes
    • Place of Presentation
      Paris,France
    • Year and Date
      2007-10-12
    • Related Report
      2007 Annual Research Report
  • [Presentation] Dynamic allosteRy in the tRyptohan-binding pRotein TRAP2007

    • Author(s)
      Heddle JG, Akashi S, Psrk SY, Tame JR
    • Organizer
      Biophysice of ligand binding to drug tagets
    • Place of Presentation
      Strasbourg,France
    • Related Report
      2007 Annual Research Report
  • [Book] 「生命秩序を担う生体超分子」蛋白質核酸酵素増刊2005

    • Author(s)
      雲財 悟, 朴 三用, Jeremy R.H.Tame
    • Total Pages
      273
    • Publisher
      共立出版
    • Related Report
      2005 Annual Research Report
  • [Publications] Heddle J, Scott DJ, Unzai S, Park SY, Tame JR.: "Crystal structures of the liganded and unliganded nickel-binding protein NikA from Escherichia coli."Journal of Biological Chemistry.. 278(50). 50322-50329 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Wada, T., Kuzuyama, T., Satoh, S., Kuramitsu, S., Yokoyama, S., Unzai, S., Tame, J.R.H., Park, S.Y.: "Crystal structure of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase, an enzyme in the non-mevalonate pathway of isoprenoid synthesis"Journal of Biological Chemistry. 278. 30022-30027 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Handa, N., et al.: "Crystal structure of the conserved protein TT1542 from Thermus thermophilus HB8."Protein Science. 12. 1621-1632 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Adachi, S., Park, S.Y., Tame, J.R.H., Shiro, Y., Shibayama, N.: "Direct observation of photolysis-induced tertiary structural changes in hemoglobin."Proc.Natl.Ac.Sci.USA. 100. 7039-7044 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Kishida, et al.: "The crystal structure and catalytic mechanism of 2-C-methyl-D-erythritol 2,4-cyclodiphosphate (MECDP) synthase, an enzyme in the nonmevalonate pathway of isoprenoid synthesis"Acta Crystallog.. D59. 23-31 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.Wada, et al.: "Structure of a conserved CoA-binding protein synthesized a cell-free system"Acta Crystallog.. D59. 1213-1218 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2018-03-28  

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