• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

p53誘導性ホスファターゼPPM1Dの機能解析用分子ツールとしての阻害剤開発

Research Project

Project/Area Number 17750148
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Chemistry related to living body
Research InstitutionHokkaido University

Principal Investigator

中馬 吉郎  Hokkaido University, 大学院・理学研究院, 助教 (40372263)

Project Period (FY) 2005 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2007: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2006: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000)
KeywordsPPM1D / ホスファターゼ / PP2C / ファージディスプレイ / ペプチド / 阻害剤 / p53 / 大腸菌発現 / 結合モチーフ
Research Abstract

1.PPM1D結合ペプチドの合成とPPMID阻害活性測定
ペプチドファージライブラリーからPM1D内部に存在するPro loop類似配列がスクリーニングされたことから、今回ペプチドP loop(21-49)をFmoc固相法により化学合成し、PPM1Dに対する阻害効果をin vitroで解析した。その結果、コントロールペプチドであるMT5ペプチドではPPM1Dペプチドはまったく阻害されないにもかかわらず、P loop(21-49)は濃度依存的にPPM1Dの活性を阻害し、IC50値594・Mを示した。今回、PPM1D内部に存在するP loopのペプチドが阻害することは、PPM1D自身による活性制御が示唆され、PPM1D阻害剤としての有用性に加え、PPM1Dの酵素活性発現メカニズムとして興味深い知見が得られた。
2.PPM1D基質を母体としたPPM1D阻害剤の開発
これまで申請者はPPM1D基質として知られている15位リン酸化p53リン酸化ペプチド(WT)の複数のアナログを用いた速度論敵解析により、PPM1D基質における酸性残基の重要性を明らかにしている。そこで、今回基質ペプチドのリン酸化セリン残基を非水解性のリン酸化セリンミミック体であるAP4に置換したペプチドアナログを作製し、PPM1Dに対する阻害活性試験を実施した。基質として最も高い親和性を示した3Eペプチドを母体としたAP4-3Eペプチドを合成しHPLC精製したところ、ジアステレオマーと考えられる2種類のペプチドが得られ、それぞれサブ・Mオーダーの高い阻害活性を示した。さらに我々は各AP4-3Eペプチドの阻害様式を解析したところ、AP4-3EペプチドがPPM1Dに対して不拮抗型阻害剤として機能しており、これまで報告されていない新規なPPM1D阻害剤であることが明らかとなった。さらに、PPM1Dのモデリング解析により、PPM1Dの活性中心近傍には正電荷に富んだ領域が複数存在しており、AP4-3Eペプチドの結合領域となることが示唆された。今回得られた基質と競合しないユニークな阻害ペプチドAP4-3EはPPM1Dを標的とした新規抗がん剤のリード化合物としてのみならず、PPM1Dの生体内における機能解明への応用が期待される。

Report

(3 results)
  • 2007 Annual Research Report
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • Research Products

    (34 results)

All 2008 2007 2006 2005

All Journal Article (17 results) (of which Peer Reviewed: 3 results) Presentation (17 results)

  • [Journal Article] Characterization of the Active Site and a Unique Uncompetitive Inhibitor of the PPM1-type Protein Phosphatase PPM1D2008

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

      Protein Pept. Lett. (In press)

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Differential Recognition of Phosphorylated Transactivation Domains of p53 by Different p300 Domains2008

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

      J. Mol. Biol. 376

      Pages: 8-12

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Correlation of Oligomeric Structure and Transcriptional Activity of p53 Mutants2008

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

      Peptide Science 2007

      Pages: 69-70

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Screening of Peptides to Stabilize the p53 Tetramer Formation from Phage Displayed Library2008

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

      Peptide Science 2007

      Pages: 315-316

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Correlation between Expression Level in E. coli and Stability of Oligomeric Structure of the p53 Tetramerization Domain2008

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

      Peptide Science 2007

      Pages: 317-318

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Structural Photomodulation of Peptides with Azobenzoate Derivative in Peptide Backbone2008

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

      Peptide Science 2007

      Pages: 447-448

    • Related Report
      2007 Annual Research Report
  • [Journal Article] Structural isoforms of the circadian neuropeptide PDF expres sed in the optic lobes of the cricket Gryllus bimaculatus. Immunocytochemical evidence from specific monoclonal antibodies2007

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

      J. Comp. Neurol. 499

      Pages: 404-421

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structural isoforms of the circadian neuropeptide PDF expressed in the optic lobes of the cricket Gryllus bimaculatus : immunocytochemical evidence from specific monoclonal antibodies2006

    • Author(s)
      Honda T.
    • Journal Title

      J. Comp. Neurol. 499

      Pages: 404-421

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Fibrillar and granular aggregation of G334V mutant p53 tetramer peptide2006

    • Author(s)
      Asanomi Y.
    • Journal Title

      Peptide Science 2005

      Pages: 397-398

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Analysis of stabilization by substitution of cyclohexylalanine for Phe341 in hydrophobic core of p53 tetramer2006

    • Author(s)
      Nomura T.
    • Journal Title

      Peptide Science 2005

      Pages: 399-402

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Characterization and application of monoclonal antibody specific for Ser(P)-Gln phosphorylation motif2006

    • Author(s)
      Iizuka K.
    • Journal Title

      Peptide Science 2005

      Pages: 499-502

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Substrate recognition of PP2C family phosphatase PPM1D2006

    • Author(s)
      Fukuda T.
    • Journal Title

      Peptide Science 2005

      Pages: 403-504

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Different structural requirements for the constitutive and the agonist-induced activities of the beta 2-adrenergic receptor2005

    • Author(s)
      Ambrosio, C
    • Journal Title

      J.Biol.Chem. 280

      Pages: 23464-23474

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification of Novel Binding Motif for Secreted Frizzled-Related Protein-12005

    • Author(s)
      Chuman, Y.
    • Journal Title

      Peptide Science 2004

      Pages: 91-94

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Triplicate Alternative Splicings that Produce Multiple Variants of the PERIOD Protein in the Silk Moth Bombym mori by Deletion/Insertion of Peptides.2005

    • Author(s)
      Takeda, Y.
    • Journal Title

      Peptide Science 2004

      Pages: 203-206

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Mechanism of amyloid-like fibrillar aggregation of mutant peptide of p53 tetramerization domain.2005

    • Author(s)
      Asanomi, Y.
    • Journal Title

      Peptide Science 2004

      Pages: 313-316

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Charactarization of superstable variant peptideof human p53 tetramerization domain.2005

    • Author(s)
      Nomura, T.
    • Journal Title

      Peptide Science 2004

      Pages: 423-426

    • Related Report
      2005 Annual Research Report
  • [Presentation] アゾベンゼンアミノ酸誘導体を導入したペプチドの光による構造制2008

    • Author(s)
      鎌田 瑠泉
    • Organizer
      日本化学会第87春季年会
    • Place of Presentation
      立教大学(東京・池袋)
    • Year and Date
      2008-03-29
    • Related Report
      2007 Annual Research Report
  • [Presentation] p53誘導性ホスファターゼPPM1Dに対する不拮抗型ペプチド性阻害2008

    • Author(s)
      八木 寛陽
    • Organizer
      日本化学会第87春季年会
    • Place of Presentation
      立教大学(東京・池袋)
    • Year and Date
      2008-03-29
    • Related Report
      2007 Annual Research Report
  • [Presentation] 非加水分解性アミドミミックを組み込んだHDAC阻害剤2008

    • Author(s)
      佐野 聡
    • Organizer
      日本化学会第87春季年会
    • Place of Presentation
      立教大学(東京・池袋)
    • Year and Date
      2008-03-29
    • Related Report
      2007 Annual Research Report
  • [Presentation] Aβアミロイドペプチドのアミノ酸付加による構造制御2008

    • Author(s)
      中馬 吉郎
    • Organizer
      日本化学会第87春季年会
    • Place of Presentation
      立教大学(東京・池袋)
    • Year and Date
      2008-03-28
    • Related Report
      2007 Annual Research Report
  • [Presentation] Stability of oligomeric structure is the key factor for transcriptional activity in tumor suppressor protein p532007

    • Author(s)
      Imagawa, T.
    • Organizer
      BMB2007:第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会
    • Place of Presentation
      パシフィコ横浜、他(神奈川県横浜市)
    • Year and Date
      2007-12-11
    • Related Report
      2007 Annual Research Report
  • [Presentation] Alternative Splicing Variants of p53-inducible Protein Phosphatase PPM1D2007

    • Author(s)
      中馬 吉郎
    • Organizer
      BMB2007:第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会
    • Place of Presentation
      パシフィコ横浜、他(神奈川県横浜市)
    • Year and Date
      2007-12-11
    • Related Report
      2007 Annual Research Report
  • [Presentation] Alignment of PPM1 type protein phosphatase PPM1D and its substrate recognition2007

    • Author(s)
      八木 寛陽
    • Organizer
      BMB2007:第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会
    • Place of Presentation
      パシフィコ横浜、他(神奈川県横浜市)
    • Year and Date
      2007-12-11
    • Related Report
      2007 Annual Research Report
  • [Presentation] CORRELATION OF OLIGOMERIC STRUCTURE AND TRANSCRIPTIONAL ACTIVITY OF P53 MUTANTS2007

    • Author(s)
      Sakaguchi, K.
    • Organizer
      第44回ペプチド討論会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2007-11-07
    • Related Report
      2007 Annual Research Report
  • [Presentation] Screening of peptides to stabilize the p53 tetramer formation from phage displayed library formation from phage displayed library2007

    • Author(s)
      野村 尚生
    • Organizer
      第44回ペプチド討論会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2007-11-07
    • Related Report
      2007 Annual Research Report
  • [Presentation] CORRELATION BETWEEN EXPRESSION LEVEL IN E. COLI AND STABILITY OF OLIGOMERIC STRUCTURE OF THE p53 TETRAMERIZATION DOMAIN2007

    • Author(s)
      Miyazaki, H.
    • Organizer
      第44回ペプチド討論会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2007-11-07
    • Related Report
      2007 Annual Research Report
  • [Presentation] Structural photomodulation of peptides with azobenzoate derivative in peptide backbone2007

    • Author(s)
      鎌田 瑠泉
    • Organizer
      第44回ペプチド討論会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2007-11-07
    • Related Report
      2007 Annual Research Report
  • [Presentation] Identification of Alternative Splicing Variants of PPM1 Protein Phospha tase PPM1D2007

    • Author(s)
      中馬 吉郎
    • Organizer
      4th International Peptide Symposium in conjunction with 7th Australian Peptide Conference and 2nd Asia-Pacific International Peptide Symposium
    • Place of Presentation
      Cairns, Great Barrier Reef, Queensland, Australia
    • Year and Date
      2007-10-21
    • Related Report
      2007 Annual Research Report
  • [Presentation] Substrate Preference of p53-inducible Protein Phosphatase PPM1D2007

    • Author(s)
      八木 寛陽
    • Organizer
      4th International Peptide Symposium in conjunction with 7th Australian Peptide Conference and 2nd Asia-Pacific International Peptide Symposium
    • Place of Presentation
      Cairns, Great Barrier Reef, Queensland, Australia
    • Year and Date
      2007-10-21
    • Related Report
      2007 Annual Research Report
  • [Presentation] Aβペプチド末端へのアミノ酸付加によるアミロイド線維の構造制御2007

    • Author(s)
      浅海 裕也
    • Organizer
      日本化学会北海道支部2007年夏季研究発表会
    • Place of Presentation
      旭川工業高等専門学校(北海道旭川市)
    • Year and Date
      2007-07-21
    • Related Report
      2007 Annual Research Report
  • [Presentation] PP2CタイプホスファターゼPPM1Dのアライメント解析と基質認識2007

    • Author(s)
      八木 寛陽
    • Organizer
      日本化学会北海道支部2007年夏季研究発表会
    • Place of Presentation
      旭川工業高等専門学校(北海道旭川市)
    • Year and Date
      2007-07-21
    • Related Report
      2007 Annual Research Report
  • [Presentation] Effects of Tumor-Associated Mutations in the p53 Tetramerization Domain on Oligomerization State and Transcriptional Activit2007

    • Author(s)
      鎌田 瑠泉
    • Organizer
      20th American Peptide Society Symposium
    • Place of Presentation
      Montreal, Quebec, Canada
    • Year and Date
      2007-06-27
    • Related Report
      2007 Annual Research Report
  • [Presentation] Study of the Interaction between Amyloid Fibrils and Gold Surfaces Modified by Self-assembled Monolayers with Various Functional Groups2007

    • Author(s)
      Tsukamoto, M
    • Organizer
      Joint Symposium on Fundamental Aspects of Nanostructured Materials and Electrocatalysis
    • Place of Presentation
      北海道大学・札幌
    • Year and Date
      2007-06-23
    • Related Report
      2007 Annual Research Report

URL: 

Published: 2005-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi