Home » 研究内容

研究内容

研究内容

動脈硬化症、がん、アルツハイマー症、ウイルス感染症など現在有効な治療法や予防法が確立されていない難病のメカニズムを生化学的・分子生物学的手法により解明し、新規の治療法や診断法を開発することを目指します。
これらの疾病に関与する因子は、細胞表面の受容体によるシグナル伝達から遺伝子の転写調節にいたるまで、複数のコンプレックスにより形成される動的なネットワークにより制御されていると考えられます。
本研究室では、バキュロウイルスを用いた抗原提示法により、疾病に関与する各タンパク質因子に対するモノクローナル抗体を作製し、高感度プロテオミクスによる分析をすることによって、これまで解析が困難であったタンパク質複合体の動的な制御機構を明らかにすることによって、新たな治療法の開発につなげます。

機能性抗体の作製とデザイン

Therapeutic Antibody making, design,

バキュロウイルスディスプレイを用いて、膜タンパク質などの機能的な抗体を作製し、がんなどの難治性疾患の治療診断にもちいることのできる抗体医薬を作ります。また、高難度の構造解析に応用したり、コンピュータによるさらに高機能な抗体医薬のデザインに挑戦します。

主要メンバー：
岩成宏子・新井修・高松佑一郎
三井健一
藤谷秀章1・山下雄史1・篠田恵子1

1 先端研バイオシミュレーション
関連主要
プロジェクト：
科学研究費基盤S
JST産学共同実用化開発事業
文部科学省　次世代がんプロジェクト
厚生労働科学研究費　革新的がん医療実用化推進研究事業
最先端プロジェクト(平成25年度終了)
ターゲットタンパク質（平成23年度終了）
NEDO新機能抗体創成プロジェクト（平成22年度終了）

論文発表

  1. Arakawa T, Kobayashi-Yurugi T, Alguel Y, Iwanari H, Hatae H, Iwata M, Abe Y, Hino T, Ikeda-Suno C, Kuma H, Kang D, Murata T, Hamakubo T, Cameron AD, Kobayashi T, Hamasaki N, Iwata S.Crystal structure of the anion exchanger domain of human erythrocyte band 3.Science. 2015 Nov 6;350(6261):680-4.
  2. Yokoi N, Fukata Y, Kase D, Miyazaki T, Jaegle M, Ohkawa T, Takahashi N, Iwanari H, Mochizuki Y, Hamakubo T, Imoto K, Meijer D, Watanabe M, Fukata M.Chemical corrector treatment ameliorates increased seizure susceptibility in a mouse model of familial epilepsyNature Medicine.Published online 08 December 2014
  3. Razvina O, Jiang S, Matsubara K, Ohashi R, Hasegawa G, Aoyama T, Daigo K, Kodama T, Hamakubo T, Naito M Differential expression of pentraxin 3 in neutrophils.Exp Mol Pathol. 2014 Nov 20. pii: S0014-4800(14)00184-1.
  4. Hamakubo T, Kusano-Arai O, Iwanari H. Generation of antibodies against membrane proteins. Biochim Biophys Acta. 2014 Aug 15. pii: S1570-9639(14)00205-2.
  5. Fujiwara K, Koyama K, Suga K, Ikemura M, Saito Y, Hino A, Iwanari H, Kusano-Arai O, Mitsui K, Kasahara H, Fukayama M, Kodama T, Hamakubo T, Momose T. A (90)Y-labelled anti-ROBO1 monoclonal antibody exhibits antitumour activity against hepatocellular carcinoma xenografts during ROBO1-targeted radioimmunotherapy. EJNMMI Res. 2014 Jun 1;4:29
  6. Patel M, Kawano T, Suzuki N, Hamakubo T, Karginov AV, Kozasa T. Gα13/PDZ-RhoGEF/RhoA signaling is essential for gastrin-releasing peptide receptor-mediated colon cancer cell migration.Mol Pharmacol. 2014 Sep;86(3):252-62.
  7. Saito Y, Miyasaka T, Hatsuta H, Takahashi-Niki K, Hayashi K, Mita Y, Kusano-Arai O, Iwanari H, Ariga H, Hamakubo T, Yoshida Y, Niki E, Murayama S, Ihara Y, Noguchi N. Immunostaining of oxidized DJ-1 in human and mouse brains. J Neuropathol Exp Neurol.2014 Jul;73(7):714-28.
  8. Tanaka T, Tahara-Hanaoka S, Nabekura T, Ikeda K, Jiang S, Tsutsumi S, Inagaki T, Magoori K, Higurashi T, Takahashi H, Tachibana K, Tsurutani Y, Raza S, Anai M, Minami T, Wada Y, Yokote K, Doi T, Hamakubo T, Auwerx J, Gonzalez FJ, Nakajima A, Aburatani H, Naito M, Shibuya A, Kodama T, Sakai J. PPARβ/δ activation of CD300a controls intestinal immunity.Sci Rep.2014 Jun 24;4:5412.
  9. Maejima T, Inoue T, Kanki Y, Kohro T, Li G, Ohta Y, Kimura H, Kobayashi M, Taguchi A, Tsutsumi S, Iwanari H, Yamamoto S, Aruga H, Dong S, Stevens JF, Poh HM, Yamamoto K, Kawamura T, Mimura I, Suehiro J, Sugiyama A, Kaneki K, Shibata H, Yoshinaka Y, Doi T, Asanuma A, Tanabe S, Tanaka T, Minami T, Hamakubo T, Sakai J, Nozaki N, Aburatani H, Nangaku M, Ruan X, Tanabe H, Ruan Y, Ihara S, Endo A, Kodama T, Wada Y. Direct evidence for pitavastatin induced chromatin structure change in the KLF4 gene in endothelial cells.PLoS One.2014 May 5;9(5):e96005.
  10. Kudo S, Caaveiro JM, Goda S, Nagatoishi S, Ishii K, Matsuura T, Sudou Y, Kodama T, Hamakubo T, Tsumoto K. Identification and characterization of the X-dimer of human P-cadherin: implications for homophilic cell adhesion.Biochemistry.2014 Mar 25;53(11):1742-52.
  11. Aramaki S, Hayashi K, Kurimoto K, Ohta H, Yabuta Y, Iwanari H, Mochizuki Y, Hamakubo T, Kato Y, Shirahige K, Saitou M. A mesodermal factor, T, specifies mouse germ cell fate by directly activating germline determinants.Dev Cell.2013 Dec 9;27(5):516-29.
  12. Rusu L, Andreeva A, Visintine DJ, Kim K, Vogel SM, Stojanovic-Terpo A, Chernaya O, Liu G, Bakhshi FR, Haberichter SL, Iwanari H, Kusano-Arai O, Suzuki N, Hamakubo T, Kozasa T, Cho J, Du X, Minshall RD. G protein-dependent basal and evoked endothelial cell vWF secretion. Blood. 2013 Sep 30. [Epub ahead of print]
  13. Ramadhanti J, Huang P, Kusano-Arai O, Iwanari H, Sakihama T, Misu T, Fujihara K, Hamakubo T, Yasui M, Abe Y.　A novel monoclonal antibody against the C-terminal region of aquaporin-4. Monoclon Antib Immunodiagn Immunother. 2013 Aug;32(4):270-6.
  14. Chow CR, Suzuki N, Kawamura T, Hamakubo T, Kozasa T. Modification of p115RhoGEF Ser(330) regulates its RhoGEF activity. Cell Signal. 2013 Nov;25(11):2085-92. doi: 10.1016/j.cellsig.2013.06.012. Epub 2013 Jun 29.
  15. Miyazaki K, Abe Y, Iwanari H, Suzuki Y, Kikuchi T, Ito T, Kato J, Kusano-Arai O, Takahashi T, Nishiyama S, Ikeshima-Kataoka H, Tsuji S, Arimitsu T, Kato Y, Sakihama T, Toyama Y, Fujihara K, Hamakubo T, Yasui M. Establishment of monoclonal antibodies against the extracellular domain that block binding of NMO-IgG to AQP4. J Neuroimmunol. 2013 Jul 15;260(1-2):107-16.
  16. Kohno T, Urao N, Ashino T, Sudhahar V, McKinney RD, Hamakubo T, Iwanari H, Ushio-Fukai M, Fukai T. Novel Role of Copper Transport Protein Antioxidant-1 in Neointimal Formation After Vascular Injury. Arterioscler Thromb Vasc Biol. 2013 Jan 24.
  17. Yumura K, Ui M, Doi H, Hamakubo T, Kodama T, Tsumoto K, Sugiyama A.　 Mutations for decreasing the immunogenicity and maintaining the function of core streptavidin.　Protein Sci. 2013 Feb;22(2):213-21.
  18. Mochizuki Y, Ohashi R, Kawamura T, Iwanari H, Kodama T, Naito M, Hamakubo T.　Phosphatidylinositol 3-phosphatase myotubularin-related protein 6 (MTMR6) is regulated by small GTPase Rab1B in the early secretory and autophagic pathways.　J Biol Chem. 2013 Jan 11;288(2):1009-21.
  19. Kusano-Arai O, Iwanari H, Mochizuki Y, Nakata H, Kodama T, Kitoh T, Hamakubo T.　Evaluation of the asparagine synthetase level in leukemia cells by monoclonal antibodies.　Hybridoma (Larchmt). 2012 Oct;31(5):325-32.
  20. Kudo S, Caaveiro JM, Miyafusa T, Goda S, Ishii K, Matsuura T, Sudou Y, Kodama T, Hamakubo T, Tsumoto K.　Structural and thermodynamic characterization of the self-adhesive properties of human P-cadherin.　Mol Biosyst. 2012 Aug;8(8):2050-3.
  21. Hino T, Arakawa T, Iwanari H, Yurugi-Kobayashi T, Ikdeda-Suno C, Nakada-Nakura Y, Kusano-Arai O, Wevand S, Shimamura T, Nomura N, Cameron A D, Kobayashi T, Hamakubo T, Iwata S & Murata T., G-protein-coupled receptor inactivation by an allosteric inverse-agonist antibody. Nature, January 29, 2012.
  22. Mimura I, Nangaku M, Kanki Y, Tsutsumi S, Inoue T, Kohro T, Yamamoto S, Fujita T, Shimamura T, Suehiro J, Taguchi A, Kobayashi M, Tanimura K, Inagaki T, Tanaka T, Hamakubo T, Sakai J, Aburatani H, Kodama T, Wada Y.　Dynamic change of chromatin conformation in response to hypoxia enhances the expression of GLUT3 (SLC2A3) by cooperative interaction of hypoxia-inducible factor 1 and KDM3A.　Mol Cell Biol. 2012 Aug;32(15):3018-32.
  23. Miyama A, Saito Y, Yamanaka K, Hayashi K, Hamakubo T, Noguchi N. Oxidation of DJ-1 induced by 6-hydroxydopamine decreasing intracellular glutathione. PLoS One. 2011;6(11):e27883. Epub 2011 Nov 21.
  24. Hayashi I, Takatori S, Urano Y, Miyake Y, Takagi J, Sakata-Yanagimoto M, Iwanari H, Osawa S, Morohashi Y, Li T, Wong PC, Chiba S, Kodama T, Hamakubo T, Tomita T, Iwatsubo T. Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin. Oncogene. 2011 Jul 4. doi: 10.1038/onc.2011.265
  25. Kanki Y, Kohro T, Jiang S, Tsutsumi S, Mimura I, Suehiro J, Wada Y, Ohta Y, Ihara S, Iwanari H, Naito M, Hamakubo T, Aburatani H, Kodama T, Minami T. Epigenetically coordinated GATA2 binding is necessary for endothelium-specific endomucin expression. EMBO J. 2011 Jun 10;30(13):2582-95
  26. Iwanari H, Nakada-Nakura Y, Kusano-Arai O, Suzuki N, Kodama T, Sakihama T, Hamakubo T. A method of generating antibodies against exogenously administered self-antigen by manipulating CD4+CD25+ regulatory T cells. J Immunol Methods. 2011 Jun 30;369(1-2):108-14.
  27. Kunii R, Jiang S, Hasegawa G, Yamamoto T, Umezu H, Watanabe T, Tsuchida M, Hashimoto T, Hamakubo T, Kodama T, Sasai K, Naito M. The predominant expression of hepatocyte nuclear factor 4α (HNF4α) in thyroid transcription factor-1 (TTF-1)-negative pulmonary adenocarcinoma.　Histopathology. 2011 Feb;58(3):467-76.
  28. Akazawa YO, Saito Y, Hamakubo T, Masuo Y, Yoshida Y, Nishio K, Shichiri M, Miyasaka T, Iwanari H, Mochizuki Y, Kodama T, Noguchi N, Niki E. Elevation of oxidized DJ-1 in the brain and erythrocytes of Parkinson disease model animals. Neurosci Lett. 2010 Oct 15;483(3):201-5.
  29. Kuwako K, Kakumoto K, Imai T, Igarashi M, Hamakubo T, Sakakibara S, Tessier-Lavigne M, Okano HJ, Okano H. Neural RNA-binding protein Musashi1 controls midline crossing of precerebellar neurons through posttranscriptional regulation of Robo3/Rig-1 expression.　Neuron. 2010 Aug 12;67(3):407-21.
  30. Seki M, Watanabe A, Enomoto S, Kawamura T, Ito H, Kodama T, Hamakubo T, Aburatani H. Human ROBO1 is cleaved by metalloproteinases and gamma-secretase and migrates to the nucleus in cancer cells FEBS Lett. 2010 Jul 2;584(13):2909-15.
  31. Suehiro JI, Hamakubo T, Kodama T, Aird WC, Minami T. Vascular endothelial growth factor activation of endothelial cells is mediated by early growth response-3. Blood. 2010 Mar 25;115(12):2520-32.
  32. Saito Y, Hamakubo T, Yoshida Y, Ogawa Y, Hara Y, Fujimura H, Imai Y, Iwanari H, Mochizuki Y, Shichiri M, Nishio K, Kinumi T, Noguchi N, Kodama T, Niki E. Preparation and application of monoclonal antibodies against oxidized DJ-1. Significant elevation of oxidized DJ-1 in erythrocytes of early-stage Parkinson disease patients. Neurosci Lett. 2009 Nov 6;465(1):1-5.
  33. Hayashi I, Takatori S, Urano Y, Iwanari H, Isoo N, Osawa S, Fukuda MA, Kodama T, Hamakubo T, Li T, Wong PC, Tomita T, Iwatsubo T. Single chain variable fragment against nicastrin inhibits the gamma-secretase activity. J Biol Chem. 2009 Oct 9;284(41):27838-47.
  34. Minami T, Yano K, Miura M, Kobayashi M, Suehiro J, Reid PC, Hamakubo T, Ryeom S, Aird WC, Kodama T. The Down syndrome critical region gene 1 short variant promoters direct vascular bed-specific gene expression during inflammation in mice. J Clin Invest. 2009 Aug;119(8):2257-70.
  35. Noritake J, Fukata Y, Iwanaga T, Hosomi N, Tsutsumi R, Matsuda N, Tani H, Iwanari H, Mochizuki Y, Kodama T, Matsuura Y, Bredt DS, Hamakubo T, Fukata M. Mobile DHHC palmitoylating enzyme mediates activity-sensitive synaptic targeting of PSD-95. J Cell Biol. 2009 Jul 13;186(1):147-60.
  36. Wakabayashi K, Okamura M, Tsutsumi S, Nishikawa NS, Tanaka T, Sakakibara I, Kitakami J, Ihara S, Hashimoto Y, Hamakubo T, Kodama T, Aburatani H, Sakai The peroxisome proliferator-activated receptor gamma/retinoid X receptor alpha heterodimer targets the histone modification enzyme PR-Set7/Setd8 gene and regulates adipogenesis through a positive feedback loop. J.Mol Cell Biol. 2009 Jul;29(13):3544-55.
  37. Okamura M, Kudo H, Wakabayashi K, Tanaka T, Nonaka A, Uchida A, Tsutsumi S, Sakakibara I, Naito M, Osborne TF, Hamakubo T, Ito S, Aburatani H, Yanagisawa M, Kodama T, Sakai J. COUP-TFII acts downstream of Wnt/beta-catenin signal to silence PPARgamma gene expression and repress adipogenesis. Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5819-24.
  38. Saitoh R, Ohtomo T, Yamada Y, Kamada N, Nezu J, Kimura N, Funahashi S, Furugaki K, Yoshino T, Kawase Y, Kato A, Ueda O, Jishage K, Suzuki M, Fukuda R, Arai M, Iwanari H, Takahashi K, Sakihama T, Ohizumi I, Kodama T, Tsuchiya M, Hamakubo T. J Immunol Methods. 2007 Apr 30;322(1-2):104-17.

FRETを用いた生細胞イメージング

FRET imaging, cell migration

FRETプローブをもちいることにより、GPCRなど膜受容体が外界から受け取ったシグナルがどのように伝達されるかを生きた細胞でリアルタイムイメージングします。
コンピュータ画像解析によって、どのように細胞が遊走するのかを明らかにします。

主要メンバー：
増田一之
小笹徹1・鈴木信周1

1先端研シグナルダイナミクス
関連主要
プロジェクト：
がん創薬プロジェクト（富士フイルム基金）
最先端プロジェクト(平成25年度終了)
科学研究費基盤S（平成２４年度終了）

論文発表

  1. Masuda K, Itoh H, Sakihama T, Akiyama C, Takahashi K, Fukuda R, Yokomizo T, Shimizu T, Kodama T, Hamakubo T. A combinatorial G protein-coupled receptor reconstitution system on budded baculovirus. Evidence for Galpha and Galphao coupling to a human leukotriene B4 receptor. J Biol Chem. 278(27):24552-62. 2003.
  2. Motoyoshi Baba, Tohru Kozasa, Takao Hamakubo, Hiroto Kuroda, Kazuyuki Masuda, Shin Yoneya and Tatsuhiko Kodama　Soft X-ray Laser Microscopy of Lipid Rafts towards GPCR-Based Drug Discovery Using Time-Resolved FRET Spectroscopy　Pharmaceuticals 2011, 4(3), 524-550

感染症とパターン認識受容体

Inflammation, pattern recognition receptor, innate immunity.

自然免疫で重要な役割を果たしている、液性パターン認識受容体ペントラキシン３（PTX3)を中心に、感染症や炎症反応における生体防御の分子メカニズムを明らかにします。好中球細胞外トラップ（Neutrophil extracellular traps: NETs）など、まったく新しい炎症像の解析から新規の治療法の開発を目指します。また、タンパク質相互作用の研究としてもコンピューターシミュレーションなど新たな分野に挑戦していきます。

主要メンバー：
太期健二・早田敬太・高松佑一郎
岩成宏子・新井修
藤谷秀章1・山下雄史1・篠田恵子1

1 先端研バイオシミュレーション
関連主要
プロジェクト：
科学研究費基盤S
A-STEP 科学技術振興機構(平成25年終了)

論文発表

  1. Razvina O, Jiang S, Matsubara K, Ohashi R, Hasegawa G, Aoyama T, Daigo K, Kodama T, Hamakubo T, Naito M Differential expression of pentraxin 3 in neutrophils.Exp Mol Pathol. 2014 Nov 20. pii: S0014-4800(14)00184-1.
  2. Daigo K, Nakakido M, Ohashi R, Fukuda R, Matsubara K, Minami T, Yamaguchi N, Inoue K, Jiang S, Naito M, Tsumoto K, Hamakubo T. Protective effect of the long pentraxin PTX3 against histone-mediated endothelial cell cytotoxicity in sepsis
    Sci. Signal., 16 September 2014; Vol. 7, Issue 343, p. ra88
  3. Daigo K, Mantovani A, Bottazzi B. The yin-yang of long pentraxin PTX3 in inflammation and immunity Immunology Letters. 2014 Sep;161(1):38-43
  4. Daigo K, Hamakubo T. Host-protective effect of circulating pentraxin 3 (PTX3) and complex formation with neutrophil extracellular traps.Front Immunol. 2012;3:378.
  5. Daigo K,Yamaguchi N, Kawamura T, Matsubara K, Jiang S, Ohashi R, Sudou Y, Kodama T, Naito M, Inoue K, and Hamakubo T. The proteomic profile of circulating pentraxin 3 (PTX3) complex in sepsis demonstrates the interaction with azurocidin 1 and other components of neutrophil extracellular traps. Mol Cell Proteomics ,　January 25, 2012
  6. Savchenko AS, Inoue A, Ohashi R, Jiang S, Hasegawa G, Tanaka T, Hamakubo T, Kodama T, Aoyagi Y, Ushiki T, Naito M. Long pentraxin 3 (PTX3) expression and release by neutrophils in vitro and in ulcerative colitis. Pathol Int. 2011 May;61(5):290-7
  7. Savchenko A, Imamura M, Ohashi R, Jiang S, Kawasaki T, Hasegawa G, Emura I, Iwanari H, Sagara M, Tanaka T, Hamakubo T, Kodama T, Naito M. Expression of pentraxin 3 (PTX3) in human atherosclerotic lesions. J Pathol. 2008 May;215(1):48-55.
  8. Imamura M, Kawasaki T, Savchenko AS, Ohashi R, Jiang S, Miyamoto K, Ito Y, Iwanari H, Sagara M, Tanaka T, Hamakubo T, Kodama T, Uchiyama M, Naito M. Lipopolysaccharide induced expression of pentraxin 3 in human neutrophils and monocyte-derived macrophages. Cell Immunol. 2007 Aug;248(2):86-94. Epub 2007 Nov 28.
  9. Inoue K, Sugiyama A, Reid PC, Ito Y, Miyauchi K, Mukai S, Sagara M, Miyamoto K, Satoh H, Kohno I, Kurata T, Ota H, Mantovani A, Hamakubo T, Daida H, Kodama T. Establishment of a high sensitivity plasma assay for human pentraxin3 as a marker for unstable angina pectoris. Arterioscler Thromb Vasc Biol. 2007 Jan;27(1):161-7.
  10. Morikawa S, Murakami T, Yamazaki H, Izumi A, Saito Y, Hamakubo T, Kodama T.Analysis of the global RNA expression profiles of skeletal muscle cells treated with statins. J Atheroscler Thromb. 12(3):121-31. 2005.

RNAスプライシング機構

RNA processing

細胞周期とRNAのプロセッシングに関与しているタンパク質WTAP (Wilms' tumor1 -associating protein) を同定しました。
WTAPに対する特異抗体を作製し、プロテオミクスによる複合体タンパク質、結合RNA解析などの網羅的解析法を駆使して、スプライシング機構と細胞周期の制御機構を明らかにします。

主要メンバー：
堀内恵子・三井健一

関連主要
プロジェクト：
がん創薬プロジェクト（富士フイルム基金）
科学研究費基盤S
　

論文発表

  1. Horiuchi K, Kawamura T, Iwanari H, Ohashi R, Naito M, Kodama T, Hamakubo T. Identification of Wilms' tumor 1-associating protein complex and its role in alternative splicing and the cell cycle. J Biol Chem. Nov 15;288(46):33292-302. 2013.
  2. Horiuchi K, Umetani M, Minami T, Okayama H, Takada S, Yamamoto M, Aburatani H, Reid PC, Housman DE, Hamakubo T, Kodama T. Wilms' tumor 1-associating protein regulates G2/M transition through stabilization of cyclin A2 mRNA. Proc Natl Acad Sci U S A. Nov 14;103(46):17278-83. 2006
  3. Umetani M, Mataki C, Minegishi N, Yamamoto M, Hamakubo T, Kodama T. Function of GATA transcription factors in induction of endothelial vascular cell adhesion molecule-1 by tumor necrosis factor-alpha. Arterioscler Thromb Vasc Biol. 21(6):917-22. 2001.

複合体プロテオミクス

proteomics

高親和性抗体と磁気ビーズを用いて、高感度のプロテオミクスによりタンパク質複合体の構成成分を網羅的に解析します。敗血症や血管疾患などこれまで解析困難だった複雑な病態を明らかにします。

主要メンバー：
太期健二・川村猛・三井健一
関連主要
プロジェクト：
科学研究費基盤S
新学術領域研究「プラズマ医療科学の創成」
がん創薬プロジェクト（富士フイルム基金）
最先端プロジェクト(平成25年度終了)

論文発表

  1. Tanaka T, Tahara-Hanaoka S, Nabekura T, Ikeda K, Jiang S, Tsutsumi S, Inagaki T, Magoori K, Higurashi T, Takahashi H, Tachibana K, Tsurutani Y, Raza S, Anai M, Minami T, Wada Y, Yokote K, Doi T, Hamakubo T, Auwerx J, Gonzalez FJ, Nakajima A, Aburatani H, Naito M, Shibuya A, Kodama T, Sakai J. PPARβ/δ activation of CD300a controls intestinal immunity.Sci Rep.2014 Jun 24;4:5412.
  2. Horiuchi K, Kawamura T, Iwanari H, Ohashi R, Naito M, Kodama T, Hamakubo T. Identification of Wilms' tumor 1-associating protein complex and its role in alternative splicing and the cell cycle. J Biol Chem. 2013 Nov 15;288(46):33292-302.
  3. Mochizuki Y, Ohashi R, Kawamura T, Iwanari H, Kodama T, Naito M, Hamakubo T.　Phosphatidylinositol 3-phosphatase myotubularin-related protein 6 (MTMR6) is regulated by small GTPase Rab1B in the early secretory and autophagic pathways.　J Biol Chem. 2013 Jan 11;288(2):1009-21.
  4. Daigo K, Hamakubo T. Host-protective effect of circulating pentraxin 3 (PTX3) and complex formation with neutrophil extracellular traps. Front Immunol. 2012;3:378.
  5. Daigo K, Yamaguchi N, Kawamura T, Matsubara K, Jiang S, Ohashi R, Sudou Y, Kodama T, Naito M, Inoue K, Hamakubo T. The proteomic profile of circulating pentraxin 3 (PTX3) complex in sepsis demonstrates the interaction with azurocidin 1 and other components of neutrophil extracellular traps. Mol Cell Proteomics. 2012 Jun;11(6):M111.015073
  6. Suzuki T, Tsuzuku J, Hayashi A, Shiomi Y, Iwanari H, Mochizuki Y, Hamakubo T, Kodama T, Nishitani H, Masai H, Yamamoto T.　Inhibition of DNA Damage-induced Apoptosis through Cdc7-mediated Stabilization of Tob.　J Biol Chem. 2012 Nov 23;287(48):40256-65.
  7. Daigo K, Kawamura T, Ohta Y, Ohashi R, Katayose S, Tanaka T, Aburatani H, Naito M, Kodama T, Ihara S, Hamakubo T. Proteomic analysis of native hepatocyte nuclear factor-4α (HNF4α) isoforms, phosphorylation status, and interactive cofactors. J Biol Chem. 2011 Jan 7;286(1):674-86.
  8. Seki M, Watanabe A, Enomoto S, Kawamura T, Ito H, Kodama T, Hamakubo T, Aburatani H. Human ROBO1 is cleaved by metalloproteinases and gamma-secretase and migrates to the nucleus in cancer cells FEBS Lett. 2010 Jul 2;584(13):2909-15.
  9. Ohguchi H, Tanaka T, Uchida A, Magoori K, Kudo H, Kim I, Daigo K, Sakakibara I, Okamura M, Harigae H, Sasaki T, Osborne TF, Gonzalez FJ, Hamakubo T, Kodama T, Sakai J. Hepatocyte nuclear factor 4alpha contributes to thyroid hormone homeostasis by cooperatively regulating the type 1 iodothyronine deiodinase gene with GATA4 and Kruppel-like transcription factor 9. Mol Cell Biol. 2008 Jun;28(12):3917-31.
  10. Sumi K, Tanaka T, Uchida A, Magoori K, Urashima Y, Ohashi R, Ohguchi H, Okamura M, Kudo H, Daigo K, Maejima T, Kojima N, Sakakibara I, Jiang S, Hasegawa G, Kim I, Osborne TF, Naito M, Gonzalez FJ, Hamakubo T, Kodama T, Sakai J. Cooperative interaction between Hepatocyte Nuclear Factor 4{alpha} and GATA transcription factors Regulates ATP-binding cassette sterol transporters ABCG5 and ABCG8. Mol Cell Biol. 2007 Apr 2

バキュロウイルステクノロジー

Baculovirus expression system

バキュロウイルス粒子に膜蛋白質が機能的に発現することを用いて、これまで困難であったにおい受容体の再構成や糖鎖合成などの膜蛋白質酵素を用いた反応・スクリーニング系を構築し、新規機能物質を同定します。

主要メンバー：
増田一之・三井健一
関連主要
プロジェクト：
最先端プロジェクト(平成25年度終了)
科学研究費基盤S

論文発表

  1. Iida M, Yoshida T, Watari A, Yagi K, Hamakubo T, Kondoh M. A baculoviral display system to assay viral entry.Biol Pharm Bull.2013;36(11):1867-9.
  2. Hamakubo T, Kusano-Arai O, Iwanari H. Generation of antibodies against membrane proteins. Biochim Biophys Acta. 2014 Aug 15. pii: S1570-9639(14)00205-2.
  3. Mitsui K, Sakihama T, Takahashi K, Masuda K, Fukuda R, Hamana H, Sato T, Hamakubo T.　Functional reconstitution of olfactory receptor complex on baculovirus.　Chem Senses. 2012 Nov;37(9):837-47.
  4. Takahashi A, Saito Y, Kondoh M, Matsushita K, Krug SM, Suzuki H, Tsujino H, Li X, Aoyama H, Matsuhisa K, Uno T, Fromm M, Hamakubo T, Yagi K., Creation and biochemical analysis of a broad-specific claudin binder. Biomaterials. 2012 Apr;33(12):3464-74.
  5. Yamagishi H, Mochizuki Y, Hamakubo T, Obata K, Ugajin T, Sato S, Kawano Y, Minegishi Y, Karasuyama H.　Basophil-derived mouse mast cell protease 11 induces microvascular leakage and tissue edema in a mast cell-independent manner. Biochem Biophys Res Commun. 2011 Dec 2;415(4):709-13.
  6. Suzuki H, Kondoh M, Kakutani H, Yamane S, Uchida H, Hamakubo T, Yagi K., The application of an alanine-substituted mutant of the C-terminal fragment of Clostridium perfringens enterotoxin as a mucosal vaccine in mice. Biomaterials. 2012 Jan;33(1):317-24.
  7. Takahashi A, Kondoh M, Uchida H, Kakamu Y, Hamakubo T, Yagi K.　Mutated C-terminal fragments of Clostridium perfringens enterotoxin have increased affinity to claudin-4 and reversibly modulate tight junctions in vitro.　Biochem Biophys Res Commun. 2011 Jul 8;410(3):466-70.
  8. Kakutani H, Takahashi A, Kondoh M, Saito Y, Yamaura T, Sakihama T, Hamakubo T, Yagi K. A novel screening system for claudin binder using baculoviral display. PLoS One. 2011 Feb 14;6(2):e16611.
  9. Uchida H, Kondoh M, Hanada T, Takahashi A, Hamakubo T, Yagi K. Mucosal vaccination using claudin-4-targeting. Biochem Pharmacol. 2010 May 15;79(10):1437-44.
  10. Saeki R, Kondoh M, Kakutani H, Tsunoda S, Mochizuki Y, Hamakubo T, Tsutsumi Y, Horiguchi Y, Yagi K. A novel tumor-targeted therapy using a claudin-4-targeting molecule. Mol Pharmacol. 2009 Oct;76(4):918-26.
  11. Sakihama T, Sato T, Iwanari H, Kitamura T, Sakaguchi S, Kodama T, Hamakubo T. A simple detection method for low-affinity membrane protein interactions by baculoviral display. PLoS ONE. 2008;3(12):e4024.
  12. Sakihama T, Masuda K, Sato T, Doi T, Kodama T, Hamakubo T. Functional reconstitution of G protein-coupled receptor-mediated adenylyl cyclase activation by a baculoviral co-display system. J Biotechnol. 2008 May 20;135(1):28-33.
  13. Saitoh R, Ohtomo T, Yamada Y, Kamada N, Nezu J, Kimura N, Funahashi S, Furugaki K, Yoshino T, Kawase Y, Kato A, Ueda O, Jishage K, Suzuki M, Fukuda R, Arai M, Iwanari H, Takahashi K, Sakihama T, Ohizumi I, Kodama T, Tsuchiya M, Hamakubo T. J Immunol Methods. 2007 Apr 30;322(1-2):104-17.
  14. Saitoh R, Ohtomo T, Ito Y, Nezu J, Kimura N, Funahashi S, Aso Y, Ohizumi I, Kodama T, Hamakubo T, Tsuchiya M. Recovery of functional peptide transporter PepT1 in budded baculovirus fraction.
  15. Ikuo Hayashi, Yasuomi Urano, Rie Fukuda, Noriko Isoo, Tatsuhiko Kodama, Takao Hamakubo Taisuke Tomita, and Takeshi Iwatsubo. Selective reconstitution and recovery of functional gamma-secretase complex on budded baculovirus particles. J Biol Chem. Sep 3;279(36):38040-6. 2004
  16. Urano Y, Yamaguchi M, Fukuda R, Masuda K, Takahashi K, Uchiyama Y, Iwanari H, Jiang SY, Naito M, Kodama T, Hamakubo T. A novel method for viral display of ER membrane proteins on budded baculovirus. Biochem Biophys Res Commun. 308(1):191-6. 2003
  17. Masuda K, Itoh H, Sakihama T, Akiyama C, Takahashi K, Fukuda R, Yokomizo T, Shimizu T, Kodama T, Hamakubo T. A combinatorial G protein-coupled receptor reconstitution system on budded baculovirus. Evidence for Galpha and Galphao coupling to a human leukotriene B4 receptor. J Biol Chem. 278(27):24552-62. 2003.

軟X線顕微鏡開発

Soft X-ray microscopy

軟X線顕微鏡で細胞、生体組織の内部構造をナノスケールの高分解能で三次元観察する技術を開発しています。細胞をはじめとする生体組織は、軽元素で構成されているので、低エネルギーの軟X線顕微鏡が必要になります。1-3keVの軟X線では、ほとんどの軽元素の吸収コントラストが観察可能で、密度(r)の微少な違いによる僅かな吸収コントラストの違いが観察できるためです。
私たちは、このような細胞や生体組織の軟X線イメージングをラボラトリーレベルで可能にすることを目的にした研究を行っています。
現在、私たちは、2keV近傍にピークを持った白色軟X線顕微鏡により、ラベリングした細胞の観察や生体組織の観察を行っています。X線顕微鏡に最適化したラベリング技術、固定化技術の研究も重要で、これらも並行して進めています。
さらに、より自然状態に近い、水をそのまま含んでいる無処理の細胞(hydrated cell)を観察するためのWater-window(W-W) X線顕微鏡の開発にも取り組んでいます。

主要メンバー：
三好元介・早田敬太
関連主要
プロジェクト：
産学共同研究(浜松ホトニクス株式会社)
科学研究費基盤S（平成２４年度終了）
NEDO産業技術実用化助成事業(平成22年度終了)

論文発表

  1. Ohsuka S, Ohba A, Onoda S, Nakamoto K, Nakano T, Miyoshi M, Soda K, Hamakubo T. Laboratory-size three-dimensional x-ray microscope with Wolter type I mirror optics and an electron-impact water window x-ray sourceRev. Sci. Instrum.2014;85;093701
  2. Takayoshi Koike, Atsuo Jouzuka, Tomonori Nakamura, Yoshihiro Onizuka, Motosuke Miyoshi and Hidenori Mimura. Structural and electrical properties if inflamed graphite at high temperature as a new field emitter, Proc. 24th International Vacuum Nanoelectronics Conference 2011 (IVNC2011), Jul. 2011, 35-36
  3. Motoyoshi Baba, Tohru Kozasa, Takao Hamakubo, Hiroto Kuroda, Kazuyuki Masuda, Shin Yoneya and Tatsuhiko Kodama. Soft X-ray Laser Microscopy of Lipid Rafts towards GPCR-Based Drug Discovery Using Time-Resolved FRET Spectroscopy Pharmaceuticals 2011, 4(3), 524-550
  4. Motosuke Miyoshi, Takao Hamakubo, Tatsuhiko Kodama, Masatoshi Tsuchya, Atsushi Koishikawa and Nobutada Aoki. Development of the low-energy soft x-ray CT instrument for the soft material structural analysis, J.Vac.Sci.Technol. B26(6), Nov/Dec 2008, 2356-2361