分子生物医学

膜タンパク質の系統的発現とモノクローナル抗体作成から複合体解明へ

バキュロウイルスを用いる膜タンパク質発現の独自技術

ゲノム解析など網羅的な解析手段により多数の機能未知の膜タンパク質やその複合体が見出され， それらの機能を探ることが課題となっている。 コレステロール代謝の研究途上で膜タンパク質を発芽型バキュロウイルス（Budded Baculovirus; BV）上に ディスプレイできることを見出した。 このＢＶ技術を用いて，Ｇタンパク質共役型受容体複合体やアルツハイマー症のγセクレターゼ複合体などの 機能的発現に成功した。 バキュロウイルスは２００nm程度の大きさで比較的安定であることからバイオチップへの応用の 可能性について研究している。 さらにＢＶ上に発現した膜タンパク質を直接免疫し，膜タンパク質に対する機能性抗体を取得する 技術を開発している（図１）。 これまで取得が困難であった膜タンパク質や複合体を認識する抗体の作成を目指す。 これらの抗体は，癌治療や診断および動脈硬化症や膠原病，神経変性疾患やウイルス感染症など 治療法の確立していない様々な疾患の新たな治療薬の開発を目標とする。

タンパク質相互作用を解析する高感度プロテオミクスの確立

膜タンパク質が外界の刺激を細胞内に伝え，そのシグナルがシグナル伝達系を経て刺激に 反応する遺伝子を誘導する。 このような刺激応答のメカニズムを解析するには，関与する複数のタンパク質の相互作用を 検出することが重要であることがわかってきた。 しかし，これまでのプロテオミクスの手法では，細胞内に微量に存在するタンパク質の解析は 非常に困難である。 本研究室では高親和性抗体を作成して低ノイズの磁性ビーズに固定化し， 少量の細胞（１０７個）から内在性転写因子をＬＣＭＳ解析にて高感度に同定する ターゲテドプロテオミクス技術を開発している(図２)。 さらにリン酸化や硫酸化など生理機能に深く関わるタンパク質修飾の解析技術を加え， 転写調節やシグナル伝達などのダイナミックなタンパク質相互作用の解析を行なうことが目標である。

特異抗体を用いるイメージング技術の開発

癌表面抗原に対する特異性の高い抗体を放射性物質でラベルし，放射線を体外から検出することにより， 癌の浸潤や転移を調べる技術を開発する(図３)。 抗体工学を用いて抗体を改変し組織移行のよいプローブを作成する。 ポジトロン核種を抗体に付加し，担癌マウスを用いて ポジトロンＣＴ（ＰＥＴ；Ｐｏｓｉｔｒｏｎ　Ｅｍｉｓｓｉｏｎ　Ｔｏｍｏｇｒａｐｈｙ）にて癌病巣部を 検出するイムノＰＥＴ技術による次世代の医療技術の開発を行なう。

図１　膜タンパク質の機能的発現と特異抗体の作成の開発

図２　タンパク質複合体のターゲテドプロテオミクス

図３　体外イメージング

Pickup publications

原著論文

1. 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.
2. 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.
3. 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.
4. Ishimoto K, Tachibana K, Hanano I, Yamasaki D, Nakamura H, Kawai M, Urano Y, Tanaka T, Hamakubo T, Sakai J, Kodama T, Doi T.　Sterol-regulatory-element-binding protein 2 and nuclear factor Y control human farnesyl diphosphate synthase expression and affect cell proliferation in hepatoblastoma cells. Biochem J. 2010 Jul 15;429(2):347-57.
5. Kakutani H, Kondoh M, Fukasaka M, Suzuki H, Hamakubo T, Yagi K. Mucosal vaccination using claudin-4-targeting. Biomaterials. Biomaterials. 2010 Jul;31(20):5463-71.
6. Uchida H, Kondoh M, Hanada T, Takahashi A, Hamakubo T, Yagi K. A claudin-4 modulator enhances the mucosal absorption of a biologically active peptide. Biochem Pharmacol. 2010 May 15;79(10):1437-44
7. Suehiro J, 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.
8. Wada Y, Ohta Y, Xu M, Tsutsumi S, Minami T, Inoue K, Komura D, Kitakami J, Oshida N, Papantonis A, Izumi A, Kobayashi M, Meguro H, Kanki Y, Mimura I, Yamamoto K, Mataki C, Hamakubo T, Shirahige K, Aburatani H, Kimura H, Kodama T, Cook PR, Ihara S. A wave of nascent transcription on activated human genes. Proc Natl Acad Sci U S A. 2009 Oct 27;106(43):18357-61.
9. Tachibana K, Takeuchi K, Inada H, Yamasaki D, Ishimoto K, Tanaka T, Hamakubo T, Sakai J, Kodama T, Doi T.　Regulation of the human SLC25A20 expression by peroxisome proliferator-activated receptor alpha in human hepatoblastoma cells.　Biochem Biophys Res Commun. 2009 Sep 11.
10. 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.
11. 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 Aug 14.
12. 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.
13. 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.
14. Komai T, Iwanari H, Mochizuki Y, Hamakubo T, Shinkai Y.　Expression of the mouse PR domain protein Prdm8 in the developing central nervous system.　Gene Expr Patterns. 2009 Oct;9(7):503-14.
15. 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.
16. Takano K, Hasegawa G, Jiang S, Kurosaki I, Hatakeyama K, Iwanari H, Tanaka T, Hamakubo T, Kodama T, Naito M.　Immunohistochemical staining for P1 and P2 promoter-driven hepatocyte nuclear factor-4alpha may complement mucin phenotype of differentiated-type early gastric carcinoma.　Pathol Int. 2009 Jul;59(7):462-70.
17. Ishimoto K, Nakamura H, Tachibana K, Yamasaki D, Ota A, Hirano K, Tanaka T, Hamakubo T, Sakai J, Kodama T, Doi T.　Sterol-mediated regulation of human lipin 1 gene expression in hepatoblastoma cells.　J Biol Chem. 2009 Aug　14;284(33):22195-205.
18. 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 J.　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.　Mol Cell Biol. 2009 Jul;29(13):3544-55.
19. 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.
20. Kazawa T, Kawasaki T, Sakamoto A, Imamura M, Ohashi R, Jiang S, Tanaka T, Iwanari H, Hamakubo T, Sakai J, Kodama T, Naito M.　Expression of liver X receptor alpha and lipid metabolism in granulocyte-macrophage colony-stimulating factor-induced human monocyte-derived macrophage.　Pathol Int. 2009 Mar;59(3):152-60.
21. Minegishi Y, Iwanari H, Mochizuki Y, Horii T, Hoshino T, Kodama T, Hamakubo T, Gotoh N.　Prominent expression of FRS2beta protein in neural cells and its association with intracellular vesicles.　FEBS Lett. 2009 Feb 18;583(4):807-14.
22. Takegoshi S, Jiang S, Ohashi R, Savchenko AS, Iwanari H, Tanaka T, Hasegawa G, Hamakubo T, Kodama T, Naito M 　Protein expression of nuclear receptors in human and murine tissues.　Pathol Int. 2009 Feb;59(2):61-72.
23. 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.
24. Sako K, Fukuhara S, Minami T, Hamakubo T, Song H, Kodama T, Fukamizu A, Gutkind JS, Koh GY, Mochizuki N 　Angiopoietin-1 induces Kruppel-like factor 2 expression through a phosphoinositide 3-kinase/AKT-dependent activation of myocyte enhancer factor 2.　J Biol Chem. 2009 Feb 27;284(9):5592-601.
25. Suzuki N, Tsumoto K, Hajicek N, Daigo K, Tokita R, Minami S, Kodama T, Hamakubo T, Kozasa T.　Activation of leukemia-associated RhoGEF by Galpha13 with significant conformational rearrangements in the interface.　J Biol Chem. 2009 Feb 20;284(8):5000-9.
26. Ishiguro T, Sugimoto M, Kinoshita Y, Miyazaki Y, Nakano K, Tsunoda H, Sugo I, Ohizumi I, Aburatani H, Hamakubo T, Kodama T, Tsuchiya M, Yamada-Okabe H.　Anti-glypican 3 antibody as a potential antitumor agent for human liver cancer.　Cancer Res. 2008 Dec 1;68(23):9832-8.
27. Nakano K, Orita T, Nezu J, Yoshino T, Ohizumi I, Sugimoto M, Furugaki K, Kinoshita Y, Ishiguro T, Hamakubo T, Kodama T, Aburatani H, Yamada-Okabe H, Tsuchiya M.　Anti-glypican 3 antibodies cause ADCC against human hepatocellular carcinoma cells.　Biochem Biophys Res Commun. 2009 Jan 9;378(2):279-84.
28. Tachibana K, Anzai N, Ueda C, Katayama T, Yamasaki D, Kirino T, Takahashi R, Ishimoto K, Komori H, Tanaka T, Hamakubo T, Ueda Y, Arai H, Sakai J, Kodama T, Doi T.　Regulation of the human PDZK1 expression by peroxisome proliferator-activated receptor alpha.　FEBS Lett. 2008 Nov 26;582(28):3884-8.
29. Sugai M, Umezu H, Yamamoto T, Jiang S, Iwanari H, Tanaka T, Hamakubo T, Kodama T, Naito M.　Expression of hepatocyte nuclear factor 4 alpha in primary ovarian mucinous tumors.　Pathol Int. 2008 Nov;58(11):681-6.
30. Yoshitake H, Shirai Y, Mochizuki Y, Iwanari H, Tsubamoto H, Koyama K, Takamori K, Ogawa H, Hasegawa A, Kodama T, Hamakubo T, Araki Y.　Molecular diversity of TEX101, a marker glycoprotein for germ cells monitored with monoclonal antibodies: variety of the molecular characteristics according to subcellular localization within the mouse testis.　J Reprod Immunol. 2008 Oct;79(1):1-11.
31. 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.
32. 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. Epub 2008 Mar 7.
33. Yorikawa C, Takaya E, Osako Y, Tanaka R, Terasawa Y, Hamakubo T, Mochizuki Y, Iwanari H, Kodama T, Maeda T, Hitomi K, Shibata H, Maki M.　Human calpain 7/PalBH associates with a subset of ESCRT-III-related proteins in its N-terminal region and partly localizes to endocytic membrane compartments.　J Biochem. 2008 Jun;143(6):731-45.
34. 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.
35. 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.
36. Tachibana K, Katayama T, Ueda C, Sumitomo M, Tagami M, Ishimoto K, Yamasaki D, Tanaka T, Hamakubo T, Sakai J, Kodama T, Obika S, Imanishi T, Doi T.　Antisense activity of 2',4'-BNA targeted to PPAR gamma in THP-1 and HCT116 cells.　Nucleic Acids Symp Ser (Oxf). 2007;(51):441-2.
37. Sugiyama A, Wada Y, Izumi A, Kobayashi M, Kohro T, Patrick CR, Hamakubo T, Kodama T.　Transcriptional activation by hypoxia and low-density lipoprotein loading in cultured vascular smooth muscle cells.　J Atheroscler Thromb. 2007 Oct;14(5):226-34.
38. Wako K, Kawasaki T, Yamana K, Suzuki K, Jiang S, Umezu H, Nishiyama T, Takahashi K, Hamakubo T, Kodama T, Naito M.　Expression of androgen receptor through androgen-converting enzymes is associated with biological aggressiveness in prostate cancer.　J Clin Pathol. 2008 Apr;61(4):448-54.
39. Reid PC, Urano Y, Kodama T, Hamakubo T.　Alzheimer's disease: cholesterol, membrane rafts, isoprenoids and statins.　J Cell Mol Med. 2007 May-Jun;11(3):383-92. Review.
40. Yoshitake H, Takahashi M, Ishikawa H, Nojima M, Iwanari H, Watanabe A, Aburatani H, Yoshida K, Ishi K, Takamori K, Ogawa H, Hamakubo T, Kodama T, Araki Y.　Aldo-keto reductase family 1, member B10 in uterine carcinomas: a potential risk factor of recurrence after surgical therapy in cervical cancer.　Int J Gynecol Cancer. 2007 Nov-Dec;17(6):1300-6.
41. 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 Jun;27(12):4248-60.
42. 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.　Viral envelope protein gp64 transgenic mouse facilitates the generation of monoclonal antibodies against exogenous membrane proteins displayed on baculovirus.　J Immunol Methods. 2007 Apr 30;322(1-2):104-17.
43. Sakamoto A, Kawasaki T, Kazawa T, Ohashi R, Jiang S, Maejima T, Tanaka T, Iwanari H, Hamakubo T, Sakai J, Kodama T, Naito M.　Expression of liver X receptor alpha in rat fetal tissues at different developmental stages.　J Histochem Cytochem. 2007 Jun;55(6):641-9.
44. Oshima T, Kawasaki T, Ohashi R, Hasegawa G, Jiang S, Umezu H, Aoyagi Y, Iwanari H, Tanaka T, Hamakubo T, Kodama T, Naito M.　Downregulated P1 promoter-driven hepatocyte nuclear factor-4alpha expression in human colorectal carcinoma is a new prognostic factor against liver metastasis.　Pathol Int. 2007 Feb;57(2):82-90.
45. Tachibana K, Anzai N, Ueda C, Katayama T, Kirino T, Takahashi R, Yamasaki D, Ishimoto K, Tanaka T, Hamakubo T, Ueda Y, Arai H, Sakai J, Kodama T, Doi T.　Analysis of PPAR alpha function in human kidney cell line using siRNA.　Nucleic Acids Symp Ser (Oxf). 2006;(50):257-8.
46. 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.
47. 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. 2006 Nov 14;103(46):17278-83. Epub 2006 Nov 6.
48. Inoue K, Kobayashi M, Yano K, Miura M, Izumi A, Mataki C, Doi T, Hamakubo T, Reid PC, Hume DA, Yoshida M, Aird WC, Kodama T, Minami T.　Histone deacetylase inhibitor reduces monocyte adhesion to endothelium through the suppression of vascular cell adhesion molecule-1 expression.　Arterioscler Thromb Vasc Biol. 2006 Dec;26(12):2652-9.
49. Ishimoto K, Tachibana K, Sumitomo M, Omote S, Hanano I, Yamasaki D, Watanabe Y, Tanaka T, Hamakubo T, Sakai J, Kodama T, Doi T.　Identification of human low-density lipoprotein receptor as a novel target gene regulated by liver X receptor alpha.　FEBS Lett. 2006 Sep 4;580(20):4929-33. Epub 2006 Aug 14.
50. Ito H, Funahashi S, Yamauchi N, Shibahara J, Midorikawa Y, Kawai S, Kinoshita Y, Watanabe A, Hippo Y, Ohtomo T, Iwanari H, Nakajima A, Makuuchi M, Fukayama M, Hirata Y, Hamakubo T, Kodama T, Tsuchiya M, Aburatani H.　Identification of ROBO1 as a novel hepatocellular carcinoma antigen and a potential therapeutic and diagnostic target.　Clin Cancer Res. 2006 Jun 1;12(11 Pt 1):3257-64.
51. Ogura T, Mio K, Hayashi I, Miyashita H, Fukuda R, Kopan R, Kodama T, Hamakubo T, Iwatsubo T, Tomita T, Sato C.　Three-dimensional structure of the gamma-secretase complex.　Biochem Biophys Res Commun. 2006 May 5;343(2):525-34.
52. Daigo K, Sugita S, Mochizuki Y, Iwanari H, Hiraishi K, Miyano K, Kodama T, Hamakubo T.　A simple hybridoma screening method for high-affinity monoclonal antibodies using the signal ratio obtained from time-resolved fluorescence assay.　Anal Biochem. 2006 Apr 15;351(2):219-28.
53. Tanaka T, Jiang S, Hotta H, Takano K, Iwanari H, Sumi K, Daigo K, Ohashi R, Sugai M, Ikegame C, Umezu H, Hirayama Y, Midorikawa Y, Hippo Y, Watanabe A, Uchiyama Y, Hasegawa G, Reid P, Aburatani H, Hamakubo T, Sakai J, Naito M, Kodama T. Dysregulated expression of P1 and P2 promoter-driven hepatocyte nuclear factor-4alpha in the pathogenesis of human cancer.
    J Pathol. 2006 Jan 6;
54. 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. Protein Expr Purif., Sep 26, 2005.
55. Tachibana K, Kobayashi Y, Tanaka T, Tagami M, Sugiyama A, Katayama T, Ueda C, Yamasaki D, Ishimoto K, Sumitomo M, Uchiyama Y, Kohro T, Sakai J, Hamakubo T, Kodama T, Doi T. Gene expression profiling of potential peroxisome proliferators activated receptor (PPAR) target genes in human hepatoblastoma cell lines inducibly expressing PPAR isoform. Nucl Recept. Oct 3;3(1):3, 2005.
56. 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.
57. Akiyama C, Shinozaki-Narikawa N, Kitazawa T, Hamakubo T, Kodama T, Shibasaki Y. Phosphatidylinositol-4-phosphate 5-kinase gamma is associated with cell-cell junction in A431 epithelial cells. Cell Biol Int. Jul;29(7):514-20. 2005.
58. Tachibana M, Ueda J, Fukuda M, Takeda N, Ohta T, Iwanari H, Sakihama T, Kodama T, Hamakubo T, Shinkai Y. Histone methyltransferases G9a and GLP form heteromeric complexes and are both crucial for methylation of euchromatin at H3-K9. Genes Dev. Apr 1;19(7):815-26. 2005.
59. Urano Y, Hayashi I, Isoo N, Reid PC, Shibasaki Y, Noguchi N, Tomita T, Iwatsubo T, Hamakubo T, Kodama T. Association of active gamma -secretase complex with lipid rafts. J Lipid Res. May;46(5):904-12. 2005.
60. Watanabe Y, Jiang S, Takabe W, Ohashi R, Tanaka T, Uchiyama Y, Katsumi K, Iwanari H, Noguchi N, Naito M, Hamakubo T, Kodama T. Expression of the LXR{alpha} Protein in Human Atherosclerotic Lesions. Arterioscler Thromb Vasc Biol. Mar;25(3):622-7. 2005.
61. Minami T, Horiuchi K, Miura M, Abid MR, Takabe W, Noguchi N, Kohro T, Ge X, Aburatani H, Hamakubo T, Kodama T, Aird WC. Vascular endothelial growth factor- and thrombin-induced termination factor, down syndrome critical region-1, attenuates endothelial cell proliferation and angiogenesis. J Biol Chem. Nov 26;279(48):50537-54. 2004.
62. 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.
63. Kohro T, Tanaka T, Murakami T, Wada Y, Aburatani H, Hamakubo T, Kodama T. A Comparison of Differences in the Gene Expression Profiles of Phorbol 12-myristate 13-acetate Differentiated THP-1 Cells and Human Monocyte-derived Macrophage.
    J Atheroscler Thromb. 11(2):88-97. 2004.
64. Morikawa S, Takabe W, Mataki C, Wada Y, Izumi A, Saito Y, Hamakubo T, Kodama T. Global analysis of RNA expression profile in human vascular cells treated with statins. J Atheroscler Thromb.;11(2):62-72. 2004.
65. Hippo Y, Watanabe K, Watanabe A, Midorikawa Y, Yamamoto S, Ihara S, Tokita S, Iwanari H, Ito Y, Nakano K, Nezu J, Tsunoda H, Yoshino T, Ohizumi I, Tsuchiya M, Ohnishi S, Makuuchi M, Hamakubo T, Kodama T, Aburatani H. Identification of soluble NH2-terminal fragment of glypican-3 as a serological marker for early-stage hepatocellular carcinoma. Cancer Res. Apr 1;64(7):2418-23. 2004.
66. Minami T, Murakami T, Horiuchi K, Miura M, Noguchi T, Miyazaki J, Hamakubo T, Aird WC, Kodama T. Interaction between Hex and GATA Transcription Factors in Vascular Endothelial Cells Inhibits flk-1/KDR-mediated Vascular Endothelial Growth Factor Signaling. J Biol Chem. May 14;279(20):20626-35. 2004.
67. Kawahara N, Wang Y, Mukasa A, Furuya K, Shimizu T, Hamakubo T, Aburatani H, Kodama T, Kirino T. Genome-wide gene expression analysis for induced ischemic tolerance and delayed neuronal death following transient global ischemia in rats.
    J Cereb Blood Flow Metab. 24(2):212-23. 2004
68. Tanaka T, Yamamoto J, Iwasaki S, Asaba H, Hamura H, Ikeda Y, Watanabe M, Magoori K, Ioka RX, Tachibana K, Watanabe Y, Uchiyama Y, Sumi K, Iguchi H, Ito S, Doi T, Hamakubo T, Naito M, Auwerx J, Yanagisawa M, Kodama T, Sakai J. Activation of peroxisome proliferator-activated receptor delta induces fatty acid beta-oxidation in skeletal muscle and attenuates metabolic syndrome.
    Proc Natl Acad Sci U S A. 100(26):15924-9. 2003.
69. Matsui Y, Saiura A, Sugawara Y, Sata M, Naruse K, Yagita H, Kohro T, Mataki C, Izumi A, Yamaguchi T, Minami T, Sakihama T, Ihara S, Aburatani H, Hamakubo T, Kodama T, Makuuchi M. Identificaton of gene expression profile in tolerizing murine cardiac allograft by co-stimulatory blockade. Physiol Genomics. 15(3):199-208. 2003.
70. Jiang S, Tanaka T, Iwanari H, Hotta H, Yamashita H, Kumakura J, Watanabe Y, Uchiyama Y, Aburatani H, Hamakubo T, Kodama T, Naito M. Expression and localization of P1 promoter-driven hepatocyte nuclear factor-4alpha (HNF4alpha) isoforms in human and rats.
    Nucl Recept. Aug 8;1(1):5. 2003.
71. Watanabe Y, Tanaka T, Uchiyama Y, Takeno T, Izumi A, Yamashita H, Kumakura J, Iwanari H, Shu-Ying J, Naito M, Mangelsdorf DJ, Hamakubo T, Kodama T. Establishment of a monoclonal antibody for human LXRalpha: Detection of LXRalpha protein expression in human macrophages.
    Nucl Recept. May 9;1(1):1. 2003
72. 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
73. 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.
74. Saiura A, Sugawara Y, Harihara Y, Sata M, Hamakubo T, Kodama T, Makuuchi M. Gene expression profile during acute rejection in rat-to-mouse concordant cardiac xenograft by means of DNA microarray. Transpl Int.;15(11):535-40. 2002.
75. Tanaka T, Takeno T, Watanabe Y, Uchiyama Y, Murakami T, Yamashita H, Suzuki A, Aoi R, Iwanari H, Jiang SY, Naito M, Tachibana K, Doi T, Shulman AI, Mangelsdorf DJ, Reiter R, Auwerx J, Hamakubo T, Kodama T. The generation of monoclonal antibodies against human peroxisome proliferator-activated receptors (PPARs). J Atheroscler Thromb. 9(5):233-42. 2002.
76. Wada H, Hashimoto K, Wada Y, Kobayashi M, Izumi A, Sugiyama A, Kohro T, Hamakubo T, Kodama T. Extensive oligonucleotide microarray transcriptome analysis of the rat cerebral artery and arachnoid tissue. J Atheroscler Thromb.; 9(5):224-32. 2002.
77. Wada Y, Sugiyama A, Yamamoto T, Naito M, Noguchi N, Yokoyama S, Tsujita M, Kawabe Y, Kobayashi M, Izumi A, Kohro T, Tanaka T, Taniguchi H, Koyama H, Hirano K, Yamashita S, Matsuzawa Y, Niki E, Hamakubo T, Kodama T. Lipid accumulation in smooth muscle cells under LDL loading is independent of LDL receptor pathway and enhanced by hypoxic conditions. Arterioscler Thromb Vasc Biol. 22(10):1712-9. 2002.
78. Morikawa S, Takabe W, Mataki C, Kanke T, Itoh T, Wada Y, Izumi A, Saito Y, Hamakubo T, Kodama T. The effect of statins on mRNA levels of genes related to inflammation, coagulation, and vascular constriction in HUVEC. Human umbilical vein endothelial cells. J Atheroscler Thromb. 9(4):178-83. 2002
79. Uchiyama Y, Huang Y, Kanamori H, Uchida M, Doi T, Takamizawa A, Hamakubo T, Kodama T. Measurement of HCV RdRp activity with C-terminal 21 aa truncated NS5b protein: optimization of assay conditions. Hepatol Res. 23(2):90-97. 2002
80. Saiura A, Kohro T, Yamamoto T, Izumi A, Wada Y, Aburatani H, Sugawara Y, Hamakubo T, Taniguchi T, Naito M, Kodama T, Makuuchi M. Detection of an up-regulation of a group of chemokine genes in murine cardiac allograft in the absence of interferon-gamma by means of DNA microarray. Transplantation. 15;73(9):1480-6. 2002.
81. Inoue K, Cynshi O, Kawabe Y, Nakamura M, Miyauchi K, Kimura T, Daida H, Hamakubo T, Yamaguchi H, Kodama T. Effect of BO-653 and probucol on c-MYC and PDGF-A messenger RNA of the iliac artery after balloon denudation in cholesterol-fed rabbits. Atherosclerosis. 161(2):353-63. 2002.
82. Mashiba S, Wada Y, Takeya M, Sugiyama A, Hamakubo T, Nakamura A, Noguchi N, Niki E, Izumi A, Kobayashi M, Uchida K, Kodama T. In vivo complex formation of oxidized alpha(1)-antitrypsin and LDL. Arterioscler Thromb Vasc Biol. 21(11):1801-8. 2001.
83. Huang Y, Uchiyama Y, Fujimura T, Kanamori H, Doi T, Takamizawa A, Hamakubo T, Kodama T. A human hepatoma cell line expressing hepatitis c virus nonstructural proteins tightly regulated by tetracycline. Biochem Biophys Res Commun. 2;281(3):732-40. 2001.
84. 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.
85. Saiura A, Mataki C, Murakami T, Umetani M, Wada Y, Kohro T, Aburatani H, Harihara Y, Hamakubo T, Yamaguchi T, Hasegawa G, Naito M, Makuuchi M, Kodama T. A comparison of gene expression in murine cardiac allografts and isografts by means DNA microarray analysis. Transplantation. Jul 27;72(2):320-9. 2001.
86. Takabe W, Kodama T, Hamakubo T, Tanaka K, Suzuki T, Aburatani H, Matsukawa N, Noguchi N. Anti-atherogenic antioxidants regulate the expression and function of proteasome {alpha}-type subunits in human endothelial cells. J Biol Chem. 276(44), 40497-501, 2001.
87. Kohro T. Nakajima T. Wada Y. Sugiyama A. Ishii M. Tsutsumi S. Aburatani H. Imoto I. Inazawa J. Hamakubo T. Kodama T. and Emi M. Genomic Structure and Mapping of Human Orphan Receptor LXR Alpha: Upreglation of LXRa mRNA During Monocyte to Macrophage Differentiation. Journal of Atherosclerosis and Thrombosis, 7(3), 145-151. 2000.
88. Morikawa S. Umetani M. Nakagawa S. Yamazaki H. Suganami H. Inoue K. Kitahara M. Hamakubo T., Kodama T. and Saito Y. Relative Induction of mRNA for HMG CoA Reductase and LDL Receptor by five Different HMG CoA Reductase Inhibitors in Cultured Human Cells. Journal of Atherosclerosis and Thrombosis, 7(3), 138-144. 2000
89. Mataki C. Murakami T. Umetani M. Wada Y. Ishii M. Tsutsumi S. Aburatani H. Hamakubo T. and Kodama T. A Novel Zinc Finger Protein mRNA in Human Umbilical Vein Endothelial Cells is Profoundly Induced by Tumor Necrosis Factor a. Journal of Atherosclerosis and Thrombosis, 7(2), 97-103. 2000.
90. Murakami T. Mataki C. Nagao C. Umetani M. Wada Y. Ishii M. Tsutsumi S. Kohro T. Saiura A. Aburatani H. Hamakubo T. and Kodama T. The Gene Expression Profile of Human Umbilical Vein Endothelial Cells Stimulated by Tumor Necrosis Factor Using DNA Microarray Analysis. Journal of Atherosclerosis and Thrombosis, 7(1), 39-44. 2000.
91. Nakajima T, Iwaki K, Hamakubo T, Kodama T, Emi M: Genomic structure of the gene encoding human 3-hydroxy-3-methyl-glutaryl coenzyme A reductase: comparison of exon/intron organization of sterol-sensing domains among four related genes. J Hum Genet, 45(5):284-9. 2000.
92. Umetani M, Nakao H, Doi T, Iwasaki A, Ohtaka M, Nagoya T, Mataki C, Hamakubo T, Kodama T. A Novel Cell Adhesion Inhibitor, K-7174, Reduces the Endothelial VCAM-1 Induction by Inflammatory Cytokines, Acting through the Regulation of GATA. Biochem Biophys Res Commun. Jun 7;272(2):370-374. 2000.
93. Shirai H, Murakami T, Yamada Y, Doi T, Hamakubo T, Kodama Structure and function of type I and II macrophage scavenger receptors. TMech Ageing Dev 1999 ;111:107-21
94. Heigoro Shirai, Mineko Yamaguchi, Ryuichiro Sato, Yoshiki Kawabe, Rie Fukuda, Tatsuhiko Kodama and Takao Hamakubo,. Degradation Enzymes for Sterol Regulatory element Binding Protein; Common Diseases:Genetic and Pathogenic Aspects of Multifunction Diseases (1999),pp131-138. Elsevier Science
95. Nakajima T., Hamakubo T., Kodama T., Inazawa J., and M. Emi. : Genomic structure and chromosomal mapping of the human sterol regulatory element binding protein (SREBP) cleavage-activating protein (SCAP) gene. J Hum Genet , 44, pp 402 - 7, 1999.
96. Takaku M, Wada Y, Jinnouchi K, Takeya M, Takahashi K, Usuda H, Naito M, Kurihara H, Yazaki Y, Kumazawa Y, Okimoto Y, Umetani M, Noguchi N, Niki E, Hamakubo T, Kodama T．An In vitro coculture model of transmigrant monocytes and foam cell formation.Arterioscler Thromb Vasc Biol Oct;19(10):2330-9, 1999.
97. Yamaguchi, M., Shirai, H., Sato, R., Kawabe,Y., Fukuda, R., Kodama, T., and Hamakubo, T. Characterization of cleaving enzymes for sterol regulatory element binding protein in hamster liver microsomes. Biochem Biophys Res Commun. 258, 542-7, 1999.
98. Kawabe Y., Suzuki T., Hayashi M., Hamakubo T., Sato R., Kodama T.: The physiological role of sterol regulatory element-binding protein-2 in cultured human cells.　Biochimica et Biophysica Acta, 1436, pp307-318, 1999.
99. Yamada Y, Doi T., Hamakubo T. and T. Kodama："Scavenger receptor family proteins: roles for atherosclerosis, host defence and disorders of the central nervous system. Cellular and Molecular Life Sciences, 54, pp628-640, 1998
100. Mosqueda-Garcia, R., Appalsamy M, Fernandez-Violante, R., Hamakubo, T., and R. Stainback. "Modulatory Effects of Endothelin on Baroreflex Activation in the Nucleus of the Solitary Tract" Europian Journal of Pharmacology, 351, pp203-207, 1998
101. Yasukiyo Mori, Hiroaki Matsubara, Satoshi Murasawa, Kazuhisa Kijima, Katsuya, Maruyama, Hiroyasu Tsukaguchi, Naohiko Okubo, Takao Hamakubo, Tadashi Inagami, Toshiji Iwasaka, Mitsuo Inada.： "Translational Regulation of Angiotensin II Type 1A Receptor Role of Upstream AUG Triplets" Hypertension, 28 : 810-817 ,1996
102. Mosqueda-Garcia, R., Fernandez-Violante, R., Hamakubo, T., and R. Stainback. "Vasopressin mediates the pressor effects of endothelin in the subfornical organ of the rat" Journal of Pharmacology & Experimental Therapeutics, 277(2):1034-1-42, 1996.
103. Mosqueda-Garcia, R., Stainback, R., Fernandez-Violante, R., and T. Hamakubo "Cardiovascular effects of endothelin in brain nuclei regulating vasopressin release." J Cardiovasc. Pharmacol., 26(suppl 3): S159-S162, 1995.
104. Kenji Ohyama, Yamano, Y., Sano, T., Nakagomi, Y., Hamakubo, T., Morishima, I., and T. Inagami："Disulfide bridges in extracellular domains of angiotensin II receptor type IA" Regulatory Peptides, 57. 141-147. 1995.
105. Yamano, Y., Ohyama, K., Kikyo, M., Sano, T., Nakagomi, Y., Inoue, Y., Nakamura, I., Guo, D-F., Hamakubo, T., and T. Inagami.："Mutagenesis and the Molecular Modeling of the Rat Angiotensin II Receptor (AT1)." Journal of Biological Chemistry, 270. pp14024-14030. 1995.
106. Yoshida, M., Hamakubo, T., and T. Inagami.；Angiotensin II-mediated Pressor Effect of Rat　Joining Peptide. American Journal of Physiology (Regulatory Integrative Comperative 　Physiology. 35), 266, R802-R808. 1994
107. Kambayashi, Y., Bardhan, S., Takahashi, K., Tsuzuki, S., Inui, H., Hamakubo, T., and T. Inagami.；Molecular Cloning of a Novel Angiotensin II Receptor Isoform Involved in Phosphotyrosine Phosphatase Inhibition. Journal of Biological Chemistry, 268; 24543-24546, 1993
108. Hamakubo, T., Yoshida, M., Nakajima, K., Watanabe, T. X., Mosqueda-Garcia, and T. Inagami. Central Cardiovascular Effects of Joining Peptide in the Genetically Hypertensive Rat.　American Journal of Physiology (Regulatory Integrative Comperative Physiology, 34), 265; R1184-R1190, 1993
109. Hamakubo, T., Furuta, H., Ichimura, M., Appalsamy, M., Mosqueda-Garcia, R., Robertson, D. and T. Inagami.；A Na Pump Inhibitor from Bovine Posterior Pituitary: Purification, Structure Determination and Its Cardiovascular effect in rat. Biochemical and Biophysical Research Communications, 189, 691-696; 1992
110. Shimizu, K., Hamamoto, T., Hamakubo, T., Lee, W. J., Suzuki, K., Nakagawa, Y., Murachi T., and T. Yamamuro.；Immunohistochemical and Biochemical Demonstration of Calcium-Dependent Cystein Proteinase (Calpain) in Calcifying Cartilage of Rats. Journal of Orthopaedic Research, 9; 26-36, 1991
111. Suzuki, K., Shimizu, K., Hamamoto, T., Nakagawa, Y., Hamakubo, T. and T. Yamamuro. Biochemical Demonstration of Calpains and Calpastatin in Osteoarthritic Synovial Fluid. Arthritis and Rheumatism, 33; 728-732, 1990
112. Kinoshita, M., Hamakubo, T., Fukui, I., Murachi, T., and H. Toyohara.；Significant Amount of Multicatalytic Proteinase Identified on Membrane from Human Erythrocyte. Journal of Biochemistry, 107; 440-444, 1990
113. Hamakubo, T., Ueda, M., Takano, E., and T. Murachi.；Distributional and Developmental Variations of Multiple Forms of Calpastatin in Mouse Brain. Journal of Enzyme Inhibition, 3; 203-210, 1990
114. Takano, E., Hamakubo, T., Kawatani, Y., Ueda, M., Kannagi, R., and T. Murachi. Multiple Forms of Calpastatin in Pig Brain. Biochemistry International, 19; 633-643. 1989
115. Murachi, T., Murakami, T., Ueda, M., Fukui, I., Hamakubo, T., Adachi, Y., and M. Hatanaka.; "The Calpain-Calpastatin System in Hematopoietic Cells" in Calcium Protein Signaling. (Hidaka, H., Carafoli, E., Means, A. R., and T. Tanaka, eds.), pp445-454. Plenum Publishing Corporation 1989
116. Ogata, N., Yonekawa, Y., Taki, W., Kannagi, R., Murachi, T., Hamakubo, T., and H. Kikuchi. Degradation of Neurofilament Protein in Cerebral Ischemia. Journal of Neurosurgery, 70; 103-107, 1989
117. Murakami, T., Ueda, M., Hamakubo, T., and T. Murachi.;；Identification of Both Calpains I and II in Nucleated Chicken Erythrocytes. Journal of Biochemistry, 103; 168-171, 1988
118. Fukui, I., Toyohara, H., Ito, K., Hamakubo, T., and T. Murachi.；Molecular and Catalytic Characterization of Intact Heterodimeric and Derived Monomeric Calpains Isolated under Different Conditions from Pig Polymorphonuclear Leukocytes. Biochemistry, 27; 3260-3267, 1988
119. Murachi, T., Hatanaka, M., and T. Hamakubo. Calpains and Neuropeptide Metabolism. in Neuropeptides and Their Peptidases. (Turner, A. J., ed.), pp202-228.Ellis-Horwood, London. 1987
120. Hamakubo, T., Ueda, M., Fucharoen, S., Wilairat, P., Pimainork, Y., and T. Murachi. Thalassemic Red Blood Cells and the Study of Membrane Protein. Contemporary Themes in Biochemistry, ICSU Short Reports, 6; 524-525, 1986
121. Hamakubo, T., Kannagi, R., Murachi, T., and A. Matus.; Distribution of Calpain in Rat Brain. Journal of Neuroscience, 6; 3103-3111, 1986
122. Miyachi, Y., Yoshimura, N., Suzuki, S., Hamakubo, T., Kannagi, R., Imamura, S., and T.　Murachi.；Biochemical Demonstration and Immunohistochemical Localization of Calpain in Human Skin. Journal of Investigative Dermatology, 86; 346-349, 1986
123. Yoshimura, N., Ohtuki, H., Hamakubo, T., Kitahara, A., Kannagi, R., and T. Murachi.；Distribution of Calpain in Various Organs of Rat: An Immunohistochemical Study. Biomedical Research, 5; 419-425, 1984

著書

124. 考える血管（児玉龍彦，浜窪隆雄）：講談社，1997
125. 浜窪隆雄，児玉龍彦：プロテインチップの開発とプロテオミクス，「プロテオミクス」；伊藤隆司，谷口壽章編，167-181頁，中山書店，2000
126. 浜窪隆雄：心臓血管系，「予防医学事典」；松島綱治，酒井敏行，石川昌，稲寺秀邦編，373-37 5頁，朝倉書店, 2005