LSBM


Expression and Interaction of Membrane Proteins

	教授
	浜窪隆雄
	Professor Takao HAMAKUBO
	専門分野：	生化学
	研究内容：	膜タンパク質の解析技術の開発。膜タンパク質，核タンパク質のターゲテドプロテオミクス。バキュロウイルス発現系による膜タンパク質特異抗体の作成技術開発。抗体を用いたイメージング技術，バイオチップ技術の研究。
		■PickUp publication
		■参考文献

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

γセクレターゼ複合体とラフトとの関係

　 γセクレターゼ複合体の活性化型が神経系細胞のラフトに存在し，スタチンによってコレステロールおよびゲラニルゲラニル化依存的に可溶性画分に移行することを示した。
　アルツハイマー症のアミロイド蓄積とコレステロール代謝の関係を示唆する。

原著論文
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.

γセクレターゼ複合体をバキュロウイルス上で再構成

　 γセクレターゼ複合体がバキュロウイルス上に再構成され，活性を有していることを示した。
　 γセクレターゼの活性化機構の研究と同時に新薬や抗体のスクリーニングに使用できる。

原著論文
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.

ＥＲ膜タンパク質のバキュロウイルス上への発現手法

　 SREBP, SCAPなどのＥＲ膜タンパク質がバキュロウイルス画分に回収されることを示した。膜タンパク質の解析に使用できる可能性を示唆した。

原著論文
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

GPCRと三量体Ｇタンパク質をバキュロウイルス上で再構成

　 GPCRが三量体Ｇタンパク質とバキュロウイルス上で再構成されることを示した。シグナル解析やチップへの応用が可能であることを示した。

原著論文
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.

血管内皮細胞でのＶＣＡＭ１の誘導におけるＧＡＴＡ因子の役割

　血管内皮細胞においてＴＮＦα刺激によるＶＣＡＭ１の誘導においてＧＡＴＡ因子およびその複合体が重要な役割を担っていることを示した。

原著論文
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.

原著論文

1. 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.

2. 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.

3. 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.

4. 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.

5. 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.

6. 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.

7. 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.

8. 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.

9. 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.

10. 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.

11. 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.

12. 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.

13. 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.

14. 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

15. 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.

16. 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.

17. 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.

18. 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

19. 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

20. 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.

21. 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.

22. 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.

23. 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.

24. 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.

25. 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

26. 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

27. 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.

28. 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.

29. 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.

30. 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.

31. 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.

32. 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.

33. 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.

34. 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.

35. 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

36. 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 ?. Journal of Atherosclerosis and Thrombosis, 7(2), 97-103. 2000.

37. 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.

38. 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.

39. 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.

40. 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

41. 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

42. 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.

43. 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.

44. 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.

45. 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.

46. 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

47. 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

48. 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

49. 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.

50. 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.

51. 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.

52. 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.

53. 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

54. 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

55. 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

56. 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

57. 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

58. 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

59. 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

60. 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

61. 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

62. 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

63. 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

64. 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

65. 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

66. 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

67. 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

68. Hamakubo, T., Kannagi, R., Murachi, T., and A. Matus.; Distribution of Calpain in Rat Brain. Journal of Neuroscience, 6; 3103-3111, 1986

69. 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

70. 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

１. 考える血管（児玉龍彦，浜窪隆雄）：講談社，1997

２. 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

３. 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