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Structural and cell biological study of sphingolipid metabolic enzyme

Research Project

Project/Area Number 19570113
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Structural biochemistry
Research InstitutionTokushima Bunri University

Principal Investigator

TSUGE Hideaki  Tokushima Bunri University, 健康科学研究所, 教授 (40299342)

Co-Investigator(Kenkyū-buntansha) KATUNUMA Nobuhiko  徳島文理大学, 健康科学研究所, 教授 (50035375)
ODA Masataka  徳島文理大学, 薬学部, 助教 (00412403)
SAKURAI Jun  徳島文理大学, 薬学部, 教授 (80029800)
Project Period (FY) 2007 – 2008
Project Status Completed (Fiscal Year 2008)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2008: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywords構造生物学 / スフィンゴミエリン代謝酵素 / スフィンゴシンキナーゼ / スフィンゴミエリン代謝系酵素 / スフィンゴミエリナーゼ / スフィンゴシン1リン酸 / 巻き戻し / 結晶化 / 結晶構造解析 / 局在化
Research Abstract

スフィンゴシン1リン酸(S1P)は、シグナル伝達分子として重要な作用を持つ。このS1P を作る酵素ヒトスフィンゴシンキナーゼ(SPHK1)の結晶構造解析を最終目標に発現、結晶化を目指す。大腸菌と蚕での発現の検討の結果、His tagged SPHK1 のインクルージョンボディからの巻き戻しを行い、0.1M のアルギニン存在下で活性を持った酵素を大量に精製した。しかしオリゴマーを取る事から巻き戻しが十分とは考えられず、この酵素から結晶化を行ったが回折を得られる結晶は得られていない。また細胞内でのSPHK1 結合タンパク質の解析を行っており、新規の結果が得られつつある。

Report

(3 results)
  • 2008 Annual Research Report   Final Research Report ( PDF )
  • 2007 Annual Research Report
  • Research Products

    (51 results)

All 2009 2008 2007 Other

All Journal Article (22 results) (of which Peer Reviewed: 22 results) Presentation (24 results) Book (2 results) Remarks (3 results)

  • [Journal Article] Structural basis of the influenza A virus RNA polymerase PB2 RNA-binding domain containing the pathogenicity-determinant lysine 627 residue2009

    • Author(s)
      Tsuge H., Kuzuhara T., Kise D. et al.
    • Journal Title

      J Biol Chem 284-11

      Pages: 6855-6860

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and X-ray diffraction analysis of RNA-primer/promoter-binding domain of influenza A virus RNA-dependent RNA polymerase PB22009

    • Author(s)
      Tsuge H., Kuzuhara T., Kise D. et al.
    • Journal Title

      Acta Crystallographica sect F Struct Biol Cryst Commun 1-65

      Pages: 144-146

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis of the influenza A virus RNA polymerase PB2 RNA-binding domain containing the pathogenicity-determinant lysine 627 residue.2009

    • Author(s)
      Kuzuhara T., Tsuge H., 等
    • Journal Title

      J Biol Chem. 284-11

      Pages: 6855-6860

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and X-ray diffraction analysis of RNA-primer/promoter-binding domain of influenza A virus RNA-dependent RNA polymerase PB2.2009

    • Author(s)
      Kuzuhara T., Tsuge H., 等
    • Journal Title

      Acta Crystallographica sect F Struct Biol Cryst Commun. 1-65

      Pages: 144-146

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of the YdiC-family protein TTHB029 from Thermus thermophilus HB8:Structural relationship with peptidoglycan N-acetylglucosamine deacetylase2008

    • Author(s)
      Tsuge H., Imagawa T. et al.
    • Journal Title

      Biochem Biophys Res Commun 367-3

      Pages: 535-541

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structure of 1-aspartate oxidase from the hyperthermophilic archaeon Sulfolobus tokodaii2008

    • Author(s)
      Tsuge H., Sakuraba H., Katunuma N. et al.
    • Journal Title

      Biochim Biophys Acta 1784-3

      Pages: 563-571

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens iota-toxin2008

    • Author(s)
      Tsuge H., Nagahama M., Oda M., Sakurai J. et al.
    • Journal Title

      Proc Natl Acad Sci U.S.A. 105-21

      Pages: 7399-7404

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structure of an archaeal alanine:glyoxylate aminotransferase2008

    • Author(s)
      Tsuge H., Katunuma N., Sakuraba H. et al.
    • Journal Title

      Acta Crystallogr D Biol Crystallogr 64-6

      Pages: 696-699

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural analysis of human glutamine:fructose-6-phosphate amidotransferase, a key regulator in type 2 diabetes2008

    • Author(s)
      Tsuge H., Nakaishi Y., Bando M. et al.
    • Journal Title

      FEBS Lett 583-1

      Pages: 163-167

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Non-native alpha-helix formation in not necessary for folding of lipocalin:Comparison of burst-phase folding between tear lipocalin and beta-lactoglobulin2008

    • Author(s)
      Tsuge H., Tsukamoto S., Yamashita T. et al.
    • Journal Title

      Proteins (In press)

      Pages: 0-0

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Structural basis of actin recognition and arginine ADP-ribosylation by Clostridium perfringens iota-toxin.2008

    • Author(s)
      Tsuge H., Nagahama M., 等
    • Journal Title

      Proc Natl Acad Sci U.S.A. 105-21

      Pages: 7399-7404

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structure of an archaeal alanine : glyoxylate aminotransferase.2008

    • Author(s)
      Sakuraba H., Tsuge H., 等
    • Journal Title

      Acta Crystallogr D Biol Crystallogr. 64-6

      Pages: 696-699

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural analysis of human glutamine : fructose-6-phosphate amidotransferase, a key regulator in type 2 diabetes.2008

    • Author(s)
      Nakaishi Y., Tsuge H., 等
    • Journal Title

      FEBS Lett. 583-1

      Pages: 163-167

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Non-native alpha-helix formation in not necessary for folding of lipocalin : Comparison of burst-phase folding between tear lipocalin and beta-lactoglobulin.2008

    • Author(s)
      Tsukamoto S., Tsuge H., 等
    • Journal Title

      Proteins. (In press)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of the YdiC-family protein TTHBO29 from Thermus thermophilus HB8:Structural relationship with peptidoglycan N-acetylglucosamine deacetylase2008

    • Author(s)
      Tsuge H., Imagawa T., 等
    • Journal Title

      Biochem Biophys Res Commun 367-3

      Pages: 535-541

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structure of l-aspartate oxidase from the hyperthermophilic archaeon Sulfblobus tokodaii2008

    • Author(s)
      Tsuge H., Sakuraba H., 等
    • Journal Title

      Biochim Biophys Acta 1784-3

      Pages: 563-571

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of archaeal highly thermostable L-aspartate dehydrogenase/NAD/citrate ternary complex2007

    • Author(s)
      Tsuge H., Yoneda K. et al.
    • Journal Title

      FEBS Journal 274-16

      Pages: 4315-4325

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Sequential aldol condensation catalyzed by hyperthermophilic 2-deoxy-d-ribose-5-phosphate aldolase2007

    • Author(s)
      Tsuge H., Sakuraba H. et al.
    • Journal Title

      Appl Environ Microbiol 73-22

      Pages: 7427-7434

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Human D-amino acid oxidase: an update and review2007

    • Author(s)
      Tsuge H., Kawazoe T. et al.
    • Journal Title

      Chem Rec 7-5

      Pages: 305-315

    • Related Report
      2008 Final Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of archaeal highly thermostable L-aspartate dehydrogenase/NAD/citrate ternary complex2007

    • Author(s)
      Tsuge H., Yoneda K., 等
    • Journal Title

      FEBS Journal 274-16

      Pages: 4315-4325

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Sequential aldol condensation catalyzed by hyperthermophilic 2-deoxy-d-ribose-5-phosphate aldolase2007

    • Author(s)
      Tsuge H., Sakuraba H., 等
    • Journal Title

      Appl Environ Microbiol 73-22

      Pages: 7427-7434

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Human D-amino acid oxidase:an update and review2007

    • Author(s)
      Tsuge H., Kawazoe T., 等
    • Journal Title

      Chem Rec 7-5

      Pages: 305-315

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Presentation] 「X 線結晶構造解析と質量分析による生理活性タンパク質の構造機能相関の研究」『アルギニン特異的ADP リボシル化毒素とアクチン複合体のX線結晶構造解析』2009

    • Author(s)
      津下英明
    • Organizer
      第3 回学術フロンティアシンポジウム
    • Place of Presentation
      徳島文理大学徳島校(徳島市)
    • Year and Date
      2009-02-28
    • Related Report
      2008 Final Research Report
  • [Presentation] 「X線結晶構造解析と質量分析による生理活性タンパク質の構造機能相関の研究」『アルギニン特異的ADPリボシル化毒素とアクチン複合体のX線結晶構造解析』2009

    • Author(s)
      津下英明
    • Organizer
      第3回学術フロンティアシンポジウム
    • Place of Presentation
      徳島文理大学徳島校(徳島市)
    • Year and Date
      2009-02-28
    • Related Report
      2008 Annual Research Report
  • [Presentation] アルギニン特異的ADP リボシル化毒素とアクチン複合体のX線結晶構造解析2008

    • Author(s)
      津下英明
    • Organizer
      北海道大学生体高分子シンポジウム
    • Place of Presentation
      北海道大学(札幌市)
    • Year and Date
      2008-12-19
    • Related Report
      2008 Final Research Report
  • [Presentation] アルギニン特異的ADPリボシル化毒素とアクチン複合体のX線結晶構造解析2008

    • Author(s)
      津下英明
    • Organizer
      北海道大学生体高分子シンポジウム
    • Place of Presentation
      北海道大学(北海道)
    • Year and Date
      2008-12-19
    • Related Report
      2008 Annual Research Report
  • [Presentation] -タンパク質・ペプチド研究の現状と展望-「アルギニン特異的ADP リボシル化毒素とアクチン複合体のX線結晶構造解析」2008

    • Author(s)
      津下英明
    • Organizer
      第18 回WS フォーラム 和光純薬工業(株)福岡営業所
    • Place of Presentation
      九州大学医系地区コラボセンター(福岡市)
    • Year and Date
      2008-11-21
    • Related Report
      2008 Final Research Report
  • [Presentation] -タンパク質・ペプチド研究の現状と展望-「アルギニン特異的ADPリボシル化毒素とアクチン複合体のX線結晶構造解析」2008

    • Author(s)
      津下英明
    • Organizer
      第18回WSフォーラム和光純薬工業(株)福岡営業所
    • Place of Presentation
      九州大学医系地区コラボセンクー(福岡市)
    • Year and Date
      2008-11-21
    • Related Report
      2008 Annual Research Report
  • [Presentation] 「X 線結晶構造解析と質量分析による生理活性タンパク質の構造機能相関の研究」『複合体タンパク質X 線結晶構造から見る酵素反応機能』2008

    • Author(s)
      津下英明
    • Organizer
      徳島文理大学・健康科学研究所・私学助成研究発表会
    • Place of Presentation
      徳島文理大学 徳島校(徳島市)
    • Related Report
      2008 Final Research Report
  • [Presentation] Aeromonas sobria セリンプロテアーゼの結晶構造解析2008

    • Author(s)
      津下英明, 宇都宮敬子 等
    • Organizer
      日本生物物理学会第1 回中国四国支部大会
    • Place of Presentation
      高知大学朝倉キャンパス(高知市)
    • Related Report
      2008 Final Research Report
  • [Presentation] Novel Family of Diflavin Enzyme:Crystal Structure of Archaeal L-Proline Dehydrogenase and the Industrial Application2008

    • Author(s)
      Tsuge H.
    • Organizer
      BIT's 1^<st> Annual World Congress of ibio 2008
    • Place of Presentation
      New Century Grand Hotel Hangzhou, China
    • Related Report
      2008 Final Research Report
  • [Presentation] Aeromonas sobriaセリンプロテアーゼの結晶構造構造解析2008

    • Author(s)
      津下英明
    • Organizer
      日本生物物理学会第1回中国四国支部大会
    • Place of Presentation
      高知大学朝倉キャンパス(高知市)
    • Related Report
      2008 Annual Research Report
  • [Presentation] Novel Family of Diflavin Enzyme : Crystal Structure of Archaeal L-Proline Dehydrogenase and the Industrial Application.2008

    • Author(s)
      Tsuge H.
    • Organizer
      BIT's 1^<st> Annual World Congress of ibio 2008
    • Place of Presentation
      New Century Grand Hotel Hangzhou, China
    • Related Report
      2008 Annual Research Report
  • [Presentation] 「X線結晶構造解析と質量分析による生理活性タンパク質の構造機能相関の研究」『複合体タンパク質X線結晶構造から見る酵素反応機能』2008

    • Author(s)
      津下 英明
    • Organizer
      徳島文理大学・健康科学研究所私学助成研究発表会
    • Place of Presentation
      徳島文理大学徳島校(徳島市)
    • Related Report
      2007 Annual Research Report
  • [Presentation] ADP リボシル化酵素とフラビン酵素の構造機能相関2007

    • Author(s)
      津下英明
    • Organizer
      第410 回ビタミンB 研究協議会
    • Place of Presentation
      京大会館(京都市)
    • Year and Date
      2007-11-17
    • Related Report
      2008 Final Research Report
  • [Presentation] ADPリボシル化酵素とフラビン酵素の構造機能相関2007

    • Author(s)
      津下 英明
    • Organizer
      第410回ビタミンB研究協議会
    • Place of Presentation
      京大会館(京都市)
    • Year and Date
      2007-11-17
    • Related Report
      2007 Annual Research Report
  • [Presentation] Crystallographic Study of Human D-Amino Acid Oxidase:Structural Basis of D-DOPA Oxidation2007

    • Author(s)
      Tsuge H., Kawazoe T., Fukui K.
    • Organizer
      THE 21^<st> SYMPOSIUM OF THE PROTEIN SOCIETY
    • Place of Presentation
      Boston, Massachusetts
    • Related Report
      2008 Final Research Report
  • [Presentation] Substrate recognition of ADP-ribosylating toxins2007

    • Author(s)
      Tsuge H.
    • Organizer
      Brookhaven National Laboratory
    • Place of Presentation
      New York
    • Related Report
      2008 Final Research Report
  • [Presentation] Crystal structure of the YdiC family protein TTHB029 from Thermus thermos thermophilus HB8 revealed structural similarity with peptidoglycan deacetylase2007

    • Author(s)
      Tsuge H., Imagawa T. et al.
    • Organizer
      ISDSB2007(The 2^<nd> International Symposium on Diffraction Structure Biology 2007)
    • Place of Presentation
      Tower Hall Funabori, Tokyo, Japan
    • Related Report
      2008 Final Research Report
  • [Presentation] Crystal structure of Aeromonas sobria serine protease, prototype kexin/furin protease:the extra occluding domain regulates protein substarte specificity2007

    • Author(s)
      Tsuge H., Kobayashi H. et al.
    • Organizer
      5^<th> General Meeting of the International Proteolysis Society(IPS2007)
    • Place of Presentation
      Patras, Greece
    • Related Report
      2008 Final Research Report
  • [Presentation] アルギニンADPリボシル化の毒素のアクチン認識分子機構2007

    • Author(s)
      津下英明, 小田真隆, 櫻井純 等
    • Organizer
      第45 回日本生物物理学会年会
    • Place of Presentation
      パシフィコ横浜会議センター(横浜市)
    • Related Report
      2008 Final Research Report
  • [Presentation] Crystallographic Study of Human D-Amino Acid OXidase:Structural Basis of D-DOPA OXidation2007

    • Author(s)
      Tsuge H., Kawazoe T., Fukui K
    • Organizer
      THE 21st SYMPOSIUM OF THE PROTEIN SOCIETY
    • Place of Presentation
      Boston,Massachusetts
    • Related Report
      2007 Annual Research Report
  • [Presentation] Substrate recognitio of ADP-ribosylating toxins2007

    • Author(s)
      Tsuge H
    • Organizer
      Brookhaven National Laboratory
    • Place of Presentation
      New York
    • Related Report
      2007 Annual Research Report
  • [Presentation] Crystal structure of the YdiC famiy protein TTHBO29 from Thermus the rmos thermophjlus HB8 revealed structural sirnilarity with peptidogly can deacetylase2007

    • Author(s)
      Tsuge H., Imagawa T., 等
    • Organizer
      ISDSB2007(The2nd International Symposium on Diffraction Structure Biology2007)
    • Place of Presentation
      Tower Hall Funabori,Tokyo,Japan
    • Related Report
      2007 Annual Research Report
  • [Presentation] Crystal structure of Aeromonas sobria serine protease,prototype kexin/furin protease:the extra occluding domain regUlates protein substarte specificity2007

    • Author(s)
      Tsuge H., Kobayashi H., 等
    • Organizer
      5th General Meeting of the International Proteolysis Society(IPS2007)
    • Place of Presentation
      Patras,Greece
    • Related Report
      2007 Annual Research Report
  • [Presentation] アルギニンADPリボシル化の毒素のアクチン認識分子機構2007

    • Author(s)
      津下 英明、小田 真隆、櫻井 純, 等
    • Organizer
      第45回日本生物物理学会年会
    • Place of Presentation
      パシフィコ横浜会議センター(横浜市)
    • Related Report
      2007 Annual Research Report
  • [Book] 構造生物学-ポストゲノム時代のタンパク質研究-(倉光成紀, 杉山政則 編)2007

    • Author(s)
      津下英明 等
    • Total Pages
      261
    • Related Report
      2008 Final Research Report
  • [Book] 構造生物学-ポストゲノム時代のタンパク質研究2007

    • Author(s)
      倉光 成紀、杉山 政則編
    • Total Pages
      261
    • Publisher
      共立出版
    • Related Report
      2007 Annual Research Report
  • [Remarks] ホームページ

    • URL

      http://inetsrv103.bunri-u.ac.jp/kenkou/

    • Related Report
      2008 Final Research Report
  • [Remarks] ホームページ

    • URL

      http://inetsrv103.bunri-u.ac.jp/kenkou/

    • Related Report
      2008 Annual Research Report
  • [Remarks]

    • URL

      http://inetsrv103.bunri-u.ac.jp/kenkou/

    • Related Report
      2007 Annual Research Report

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Published: 2007-04-01   Modified: 2016-04-21  

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