平成20年 病態システム酵素学研究部門(旧遺伝制御学部門)

1)D-アミノ酸酸化酵素の構造と病態生理学的意義の解明

統合失調症は、多因子の病因に基づく難治性精神疾患であり、D-アミノ酸酸化酵素は、本酵素活性を正に制御する新規遺伝子G72とともに、統合失調症の疾患感受性遺伝子である。脳内アストログリアにおける機能を解析した結果、本酵素が従来否定的であった大脳由来のグリアでも、遺伝子発現されていることを明らかとした。さらに、細胞外D-セリンをグリアが取り込み、積極的に代謝することを明らかとして、 NMDA受容体のコアゴニストであるD-セリンの代謝調節を介して、本酵素が統合失調症の発症とその病態に密接に関与することを提唱している。ヒト酵素蛋白のX線結晶構造解析にも成功し、統合失調症病態における酵素活性化の分子機構の解明と新規統合失調症治療薬としての阻害剤の開発を目指した製薬企業との共同研究を展開している。これらの研究成果について、2005年度は3つの国際会議(第15回国際フラビンシンポジウム、第9回国際アミノ酸会議、国際ビタミン・補酵素・バイオファクター会議)で招待講演を行っている。2006年度には、韓国生命科学会の国際シンポジウム並びに東国大学主催のシンポジウムに招待され、講演を行った。2007年度は、第3回D-アミノ酸研究会学術集会を開催し、全国から100名を越える参加者を得て、D-アミノ酸に関する学際的研究の推進に貢献するとともに、日本生化学誌での「D-アミノ酸制御システムのニューバイオロジー:Frontier Science in Amino Acid and Protein Research」特集号を企画して、研究成果の情報発信を行った。さらに2008年度には、我が国におけるビタミン学の進歩発展に功績があったとして、「D-アミノ酸代謝酵素システムの機能と構造に関する疾患酵素学的研究」に対して日本ビタミン学会賞が授与された。本酵素と同じくフラビン酵素であるL- 乳酸酸化酵素のX線結晶構造解析にも成功し,その構造機能相関を明らかとしている。


2)新規アポトーシス制御因子ヌクリングの情報伝達機構と機能の解明

ヌクリングはマウス胚性腫瘍細胞より単離された新規分子で【JB 論文賞受賞】、アポトーシス制御分子であるApaf-1/プロカスパーゼ9複合体(アポプトゾーム)と結合して、アポトーシス経路を正に制御して細胞死を誘導することを明らかとした【特許登録済】。また、アポトーシス関連分子であるガレクチン-3の発現をヌクリングが、転写因子NF-kB を介して制御することを見いだした。さらに、本遺伝子を欠損するノックアウトマウスの樹立に成功し、このマウスを用いた薬剤誘導性パーキンソン病モデルでは、ドーパミン神経細胞特異的なアポトーシスが、抑制されていることを明らかとして、新規治療戦略構築の物質的基盤を提供している。

 

1997~2008年社会貢献実績報告書

(1)特許権等の知的財産権

知的財産権の内容、発明者名、権利者、知的財産権の種類、番号、出願年月日、取得年月日を記入。

  1. 新規アポトーシス誘導タンパク質及びそれをコードする遺伝子
    発明者:福井 清、坂井隆志
    特許権者:大鵬薬品工業(株)、福井 清、坂井隆志
    特許登録済
    特許3985131号(登録年月日:2007年7月20日)

 

(2)社会的活動

国際フラビンシンポジウムの組織委員、日本生化学会の理事、科学研究費審査専門委員4回等の活動を行って、国内・国際的貢献に努めている。2005年の日本理科教育学会全国大会では、我が国の理科教育に携わる教員に対して特別講演を行った。さらに徳島県教育委員会外部講師招聘事業にて、地域における教育実践も行っている。

 

業績
  1. The Effect of Risperidone on D-Amino Acid Oxidase Activity as a Hypothesis for a Novel Mechanism of Action in the Treatment of Schizophrenia.
    Journal of Psychopharmacol., in press IF: 3.782
    Rabab M. Abou El-Magd, Hwan Ki Park, Tomoya Kawazoe, Sanae Iwana, Koji Ono, Seong Pil Chung, Motoshige Miyano, Kazuko Yorita, Takashi Sakai, Kiyoshi Fukui
  2. Nucling interacts with nuclear factor-κB, regulating its cellular distribution
    FEBS Journal, in press IF: 3.396
    Li Liu, Takashi Sakai Hoang Nam Tran, Rika Mukai-Sakai, Ryuji Kaji, Kiyoshi Fukui
  3. Positive regulation of promoter activity of human 3-phosphoglycerate dehydrogenase (PHGDH) gene is mediated by transcription factors Sp1 and NF-Y
    GENE 414 (1-2), 106-114 (2008) IF: 2.871
    Do Youn Jun, Hae Sun Park, Ji Young Lee, Joo Youn Baek, Hwan Ki Park, Kiyoshi Fukui, Young Ho Kim
  4. Chlorpromazine oligomer is a potentially active substance that inhibits human D-amino acid oxidase, product of a susceptibility gene for schizophrenia
    J. Enzym. Inhib. Med. Chem. 23(6), 901-911(2008) IF: 1.343
    Sanae Iwana, Tomoya Kawazoe, Hwan Ki Park, Koichiro Tsuchiya, Koji Ono, Kazuko Yorita, Takashi Sakai, Takenori Kusumi, Kiyoshi Fukui
  5. Human D-amino acid oxidase: an update and review
    The Chemical Record 7 (5), 305-315 (2007) IF: 2.641
    Tomoya Kawazoe, Hwan Ki Park, Sanae Iwana, Hideaki Tsuge, Kiyoshi Fukui
  6. Crystallographic study on the interaction of L-lactate oxidase with pyruvate at 1.9Å resolutionBiochem. Biophys. Res. Commun. 358 (4), 1002-1007 (2007) IF: 2.749
    Shun Jie Li, Yasufumi Umena, Kazuko Yorita, Takeshi Matsuoka, Akiko Kita, Kiyoshi Fukui, Yukio Morimoto
  7. Structural basis of D-DOPA oxidation by D-amino acid oxidase: alternative pathway for dopamine biosynthesis
    Biochem. Biophys. Res. Commun. 355 (2), 385-391 (2007) IF: 2.749
    Tomoya Kawazoe, Hideaki Tsuge, Takahito Imagawa, Kenji Aki, Seiki Kuramitsu, Kiyoshi Fukui
  8. Immunohistochemical and biochemical determination of D-serine in the cultured astrocytes and neurons from the rat neocortex
    Neurosci. Res. 58, S137 (2007) IF: 2.121
    Nagisa Kozuka, Asami Umino, Dai Shimazu, Tetsuro Kubota, Reiji Semba, Tomoya Kawazoe, Kiyoshi Fukui, Naoki Yamamoto, Toru Nishikawa
  9. Crystal structure of human D-amino acid oxidase:context-dependent variability of the backbone conformation of the VAAGL hydrophobic stretch located at the si-face of the flavin ring
    Protein Sci. 15 (12), 2708 - 2717 (2006) IF; 3.135
    Tomoya Kawazoe, Hideaki Tsuge, Mirella S. Pilone, Kiyoshi Fukui
  10. The crystal structure of L-lactate oxidase from Aerococcus viridans at 2.1Å resolution reveals the mechanism of strict substrate recognition
    Biochem. Biophys. Res. Commun. 350 (2), 249-256 (2006) IF: 2.749
    Yasufumi Umena, Kazuko Yorita, Takeshi Matsuoka, Akiko Kita, Kiyoshi Fukui, Yukio Morimoto
  11. Characterization of human D-amino acid oxidase
    FEBS Letters 580 (9), 2358-2364 (2006) IF: 3.263
    Gianluca Molla, Silvia Sacchi, Mariagazia Bernasconi, Mirella S. Pilone, Kiyoshi Fukui, Loredano Pollegioni,
  12. Attenuation of MPTP-induced neurotoxicity and locomotor dysfunction in Nucling-deficient mice via suppression of the apoptosome pathway
    J. Neurochem. 97 (4), 1126-1135 (2006) IF: 4.451
    Xichuan Teng, Takashi Sakai, Li Liu, Rika Sakai, Ryuji Kaji, Kiyoshi Fukui
  13. Potential Role for Astroglial D-Amino Acid Oxidase in Extracelluar D-Serine Metabolism and Cytotoxicity
    J. Biochem. 139 (2), 295-304 (2006) IF: 2.020
    Hwan Ki Park. Yuji Shishido, Sayaka Ichise-Shishido, Tomoya Kawazoe, Koji Ono, Sanae Iwana, Yumiko Tomita, Kazuko Yorita, Takashi Sakai, Kiyoshi Fukui
  14. Functional Roles and Pathophysiology of Brain D-Amino Acid Oxidase
    Flavins and Flavoproteins 2005, 853-860 (2005) IF: -
    Kiyoshi Fukui, Hwan Ki Park, Tomoya Kawazoe, Koji Ono, Sanae Iwana, Yumiko Tomita, Kazuko Yorita, Takashi Sakai and Young Ho Kim
  15. Purification and Crystal Structure of Human D-Amino Acid Oxidase
    Flavins and Flavoproteins 2005, 33-36 (2005) IF: -

    Tomoya Kawazoe, Sanae Iwana, Koji Ono, Hwan Ki Park, Kazuko Yorita, Yumiko Tomita, Hideaki Tsuge and Kiyoshi Fukui
  16. Molecular Basis of Schizophrenia: Characterization of Human D-Amino Acid Oxidase
    Flavins and Flavoproteins 2005,861-866(2005) IF: -
    Gianluca Molla, Mirella S. Pilone, Silvia Sacchi, Mariagrazia Barnasconi, Kiyoshi Fukui and Loredano Pollegioni
  17. Crystal Structures of Wild-type and R181M L-Lactate Oxidase from Aerococcus viridan
    Flavins and Flavoproteins 2005, 43-48 (2005) IF: -
    Kazuko Yorita, Yasufumi Umena, Takeshi Matsuoka, David P. Ballou, Makoto Abe, Akiko Kita, Tomitake Tsukihara, Yukio Morimoto and Kiyoshi Fukui
  18. H265Q L-Lactate Oxidase from Aerococcus viridans
    Flavins and Flavoproteins 2005, 37-41 (2005) IF: -
    Kazuko Yorita, Takeshi Matsuoka, David P. Ballou and Kiyoshi Fukui
  19. Astroglial D-amino acid oxidase is the key enzyme to metabolize extracellular D-serine, a neuromodulator of N-methyl-D-aspartate receptor.
    Amino Acids, 29 (1), 61-62 (2005) IF: 2.780
    Kiyoshi Fukui, Hwan Ki Park, Tomoya Kawazoe, Koji Ono, Sanae Iwana, Kazuko Yorita, Takashi Sakai
  20. Crystallization and preliminary X-ray diffraction study of L-lactate oxidase (LOX), R181M mutant, from Aerococcus viradans
    Acta Cryst F 61, 439-441 (2005) IF: 0.645
    Yasufumi Umena, Kazuko Yorita, Takeshi Matsuoka, Makoto Abe, Akiko Kita, Kiyoshi Fukui, Tomitake Tsukihara, Yukio Morimoto
  21. Nucling Recruits Apaf-1/Pro-caspase-9 Complex for the Induction of Stress-Induced Apoptosis
    J. Biol. Chem. 279(39), 41131-41140 (2004) IF: 5.581Takashi Sakai, Li Liu, Xichuan Teng, Rika Mukai-Sakai, Hidenori Shimada, Ryuji Kaji, Tasuku Mitani, Mitsuru Matsumoto, Kazunori Toida, Kazunori Ishimura, Yuji Shishido, Tak W. Mak and Kiyoshi Fukui
  22. Nucling mediates apoptosis by inhibiting expression of galectin-3 through interference with nuclear factor κB signalling
    Biochem. J. 380(1), 31-41 (2004) IF: 4.009
    Li Liu, Takashi Sakai, Nobuya Sano and Kiyoshi Fukui
  23. Identification of a Novel, Embryonal Carcinoma Cell-Associated Molecule, Nucling, That Is Up-regulated during Cardiac Muscle Differentiation
    J. Biochem. 133(4), 429-436 (2003) IF: 2.020 [JB論文賞]
    Takashi Sakai, Li Liu, Yuji Shishido and Kiyoshi Fukui
  24. Astroglial expression of D-amino acid oxidase gene
    Flavins and Flavoproteins 2002, 853-856 (2002) IF: -
    Atsuhiko Suzue, Shinji Nagahiro, Yoshiteru Urai, Osamu Jinnouchi, Tokujiro Kanamori, Kyung Tak Kwak, Hwan Ki Park, Tomoya Kawazoe, Sayaka Ichise, Yuji Shishido, Yumiko Tomita, Takashi Sakai and Kiyoshi Fukui
  25. Gene expression of D-amino Acid oxidase in cultured rat astrocytes: regional and cell type specific expression
    Neurosci. Lett. 324(2), 101-104 (2002) IF: 2.085
    Yoshiteru Urai, Atsuhiko Suzue, Osamu Jinnouchi, Kyung Tak Kwak, Shinji Nagahiro and Kiyoshi Fukui
  26. Chk2 Is a Tumor Suppressor That Regulates Apoptosis in both an Ataxia Telangiectasia Mutated (ATM)-Dependent and an ATM-Independent Manner
    Mol. Cell Biol. 22(18), 6521-6532 (2002) IF: 6.420
    Atsushi Hirao, Alison Cheung, Gordon Duncan, Pierre-Marie Girard, Andrew J. Elisa, Andrew Wakeham, Hitoshi Okada, Talin Sarkissian, Jorge A. Wong, Takashi Sakai, Elisa de Stanchina, Robert G. Bristow, Toshio Suda, Scott W. Lowe, Penny A. Jeggo, Stephen J. Elledge and Tak W. Mak
  27. Function of two arginine residues of L-lactate oxidase
    Oxygen and Life 2002, 69-75 (2002) IF: -
    Kazuko Yorita, Hideo Misaki, Mitsuru Matsumoto and Vincent Massey
  28. Positioning of Arg-268 in L-Lactate Oxidase from Aerococcus viridans
    Flavins and Flavoproteins 2002, 423-427 (2002) IF: -
    Kazuko Yorita, Toshiyuki Watanabe, Hideo Misaki and Vincent Massey
  29. Minoxidil-Induced hair growth is mediated by adenosine in cultured dermal papilla cells: possible involvement of sulfonylurea receptor 2B as a target of minoxidil
    J. Invest. Dermatol. 117(6), 1594-1600 (2001) IF: 4.829
    Ming Li, Azusa Marubayashi, Yutaka Nakaya, Kiyoshi Fukui and Seiji Arase
  30. Design and properties of human D-amino acid oxidase with covalently attached flavin
    Proc. Natl. Acad. Sci. USA 97(7), 3089-3093 (2000) IF: 9.598
    Andrei A. Raibekas, Kiyoshi Fukui and Vincent Massey
  31. Hyperlipemia and Early Pancreatic Injury Induced by Ethanol Intake in Rats
    J. Nutr. Sci. Vitamol. 46(6), 297-301 (2000) IF: 0.784
    Chie Yuasa, Osamu Miyoshi, Kiyoshi Fukui and Toshinori Oka
  32. NBP1 (Nap1 Binding Protein 1), an essential gene for G2/M transition of Saccharomyces cerevisiae, encodes a protein of distinct sub-nuclear localization
    GENE 246, 395-404 (2000) IF: 2.871
    Yoshihisa Shimizu, Tomohiro Akashi, Asuko Okuda, Akihiko Kikuchi and Kiyoshi Fukui
  33. Interaction of two arginine residues in lactate oxidase with the enzyme flavin; Conversion of FMN to 8-formyl-FMN.
    Proc. Natl. Acad. Sci. USA. 97(24), 13039-13044 (2000) IF: 9.598
    Kazuko Yorita, Takeshi Matsuoka, Hideo Misaki and Vincent Massey
  34. On the interpretaion of quantitative structure-function activity relationship data for lactate oxidase.
    Proc. Natl. Acad. Sci. USA. 97(6), 2480-2485 (2000) IF: 9.598
    Kazuko Yorita, Hideo Misaki, Bruce A. Palfey and Vincent Massey
  35. Abnormal Immune Function of Hemopoietic Cells from Alymphoplasia (aly) Mice, a Natural Strain with Mutant NF-kB-Inducing Kinase
    J. Immunol. 165(2), 804-812 (2000) IF: 6.068
    Takuji Yamada, Tasuku Mitani, Kazuko Yorita, Daisuke Uchida, Akemi Matsushima, Kisuke Iwamasa, Shigeru Fujita and Mitsuru Matsumoto
  36. Oral Administration of Sepimostat Mesilate Prevents Acute Alcohol Pancreatic Injury in Rats
    J. Pharm. Pharmacol. 51(7), 867-871 (1999) IF: 1.718
    Chie Yuasa, Kenji Irimura, Minoru Oda, Kiyoshi Fukui and Toshinori Oka
  37. Gene Expression of D-Amino Acid Oxidase in Nervous System
    Flavins and Flavoproteins 1999 887-890 (1999) IF: -
    Tokujiro Kanamori, Mariko Obayashi, Osamu Jinnouchi, Kazuya Kanda, Yoshiteru Uari, Yuji Shishido, Atsuhiko Suzue, Takashi Sakai and Kiyoshi Fukui
  38. Reaction Mechanism of L-Lactate Oxidase from Aerococcus viridan
    Flavins and Flavoproteins 1999, 479-482 (1999) IF: -
    Kazuko Yorita, Toshiyuki Watanabe, Hideo Misaki and Vincent Massey
  39. Functional Replacement of the Intracellular Region of the Notch1 Receptor by Epstein-Barr Virus Nuclear Antigen 2
    J. Virol. 72(7), 6034-6039 (1998) IF: 5.332
    Takashi Sakai, Yoshihito Taniguchi, Kumiko Tamura, Shigeru Minoguchi, Takataro Fukuhara, Lothar J. Strobl, Ursula Zimber-Strobl, George W. Bornkamm and Tasuku Honjo
  40. Differences in ethidium bromide and 4'-6-diamidino-2-phenylindole stainig profiles with regard to DNA fragmentation during apoptosis
    Biochem. Mol. Biol. Int. 46, 1055-1061 (1998) IF: -
    Masayuki Shono, Ichiro Shimizu, Toshihiro Omoya, Aya Hiasa, Hirohito Honda, Yumiko Tomita and Susumu Ito
  41. L-Lactate oxidase from Aerococcus viridans crystallized as an octamer. Preliminary X-ray studies.
    Biochimie 80(4), 309-312 (1998) IF: 2.899
    Yukio Morimoto, Kazuko Yorita, Kenji Aki, Hideo Misaki and Vincent Massey
  42. Tissue Specific Expression of the D-Amino Acid Oxidase Gene
    Flavins and Flavoproteins, 1996 199-202 (1997) IF: -
    Kiyoshi Fukui, Tokujiro Kanamori, Osamu Jinnouchi and Oded Ben-Yoseph
  43. Is There D-Amino acid oxidase in the Mouse Liver?
    Flavins and Flavoproteins 1996, 203-206 (1997) IF: -
    Ryuichi Konno, Akira Niwa, Masato Sasaki, Jumpei Enami, Setsuko Asakura and Kiyoshi Fukui
  44. Basal forebrain and cerebral cortical muscarinic receptors mediate in crease in cortical blood flow provoked by periaqueductal gray matter
    Neuroscience 79(2), 571-579 (1997) IF: 3.352
    Masatsugu Nakai, Jun Ogata, Kiyoshi Fukui, Y Nakai and Michiyuki Maeda
  45. D-Amino acid oxidase is not present in the mouse liver
    Biochim. Biophys. Acta-Gen. Subj.1335, 173-181 (1997) IF: 2.855
    Ryuichi Konno, Masato Sasaki, Setsuko Asakura, Kiyoshi Fukui, Jumpei Enami and Akira Niwa
  46. Involvement of RBP-J in biological functions of mouse Notch1 and its derivatives
    Development 124, 4133-4141 (1997) IF: 7.293Hisamune Kato, Yoshihito Taniguchi, Hisanori Kurooka, Shigeru Minoguchi, Takashi Sakai, Saeko Nomura-Okazaki, Kumiko Tamura and Tasuku Honjo
  47. On the reaction mechanism of L-lactate oxidase: Quantitative structure-activity analysis of the reaction with para-substituted L-mandelates
    Proc. Natl. Acad. Sci. USA 94(18), 9590-9595 (1997) IF: 9.598
    Kazuko Yorita, Karl Janko, Kenji Aki, Sandora Ghisla, Bruce A. Palfey and Vincent Massey
  48. Conversion of L-Lactate Oxidase to a Long Chain L-α-Hydroxyacid Oxidase by Replacement of Ala-95 to Gly
    Flavins and Flavoproteins 1996, 591-594 (1997) IF: -
    Kazuko Yorita, Kenji Aki, Toyomi Ohkuma-Soyejima, Toshio Kokubo, Hideo Misaki and Vincent Massey