脂質メディエーター結合タンパク質の網羅的同定と生理機能の解明

• Project/Area Number: 16K08596
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Juntendo University
• Principal Investigator: 奥野 利明 順天堂大学, 医学(系)研究科(研究院), 准教授 (60361466)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Granted (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 脂質 / GPCR / インバースアゴニスト / BLT1 / ロイコトリエン / 立体構造解析 / LTB4 / 脂質生化学 / ケミカルバイオロジー / 生体分子 / 生理活性

Outline of Annual Research Achievements

昨年ロイコトリエエンB4(LTB4)の受容体BLT1とインバースアゴニストBIIL260の複合体の結晶構造解析に成功した。その結果、BIIL260に含まれるベンザミジン基が、BLT1のNa+ー水クラスター結合部位に結合していることを明らかにした。ベンザミジン基が不活性状態のBLT1の立体構造を安定化していると考えられるが、BLT1の恒常活性変異体は得られていなかったため、BIIL260がインバースアゴニストであるかは否かは明らかでなかった。そこで、過去のシステイニルロイコトリエンD4受容体(CysLT2)の変異体が恒常活性を示す報告などを参考に、BLT1のヘリックス3のアミノ酸を点変異させたところ、TGFアルファ切断アッセイで恒常活性を示すことを明らかにした。このBLT1変異体を用いたTGFアルファ切断アッセイにおいて、BIIL260は容量依存的にインバースアゴニスト活性を示した。したがって、BIIL260はBLT1のインバースアゴニストであり、IC50も比較的小さいことがわかった。この結果は、ベンザミジン基がBLT1の7回膜貫通ヘリックス束の活性状態への構造変化を阻害し、Na+ー水クラスターと同様に不活性状態の立体構造の維持に寄与することを示唆している。
さらに、ベンザミジン基を有するBIIL260とは異なる構造の化合物がBLT1に対するインバースアゴニスト活性を示すことを見出しており、今後、構造活性相関の検討によって新たなBLT1拮抗薬の開発を目指す。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

BLT1に対するインバースアゴニスト活性を有する化合物を見出しており、構造活性相関の検討により、新規BLT1拮抗薬を開発する基礎データが得られているため。

Strategy for Future Research Activity

BLT1に対してより活性の高いインバースアゴニストを見出し、新規BLT1拮抗薬の開発を目指す。

Research Products

• [Journal Article] Dietary ω-3 fatty acids alter the lipid mediator profile and alleviate allergic conjunctivitis without modulating Th2 immune responses (2019)
  • Author(s): Hirakata T., Lee H. C., Ohba M., Saeki K., Okuno T., Murakami A., Matsuda A., Yokomizo T.
  • Journal Title: The FASEB Journal Volume: 33 Pages: 3392-3403
  • DOI: 10.1096/fj.201801805r
  • Peer Reviewed / Open Access
• [Journal Article] Na(+)-mimicking ligands stabilize the inactive state of leukotriene B4 receptor BLT1. (2018)
  • Author(s): Hori Tetsuya、Okuno Toshiaki、Hirata Kunio、Yamashita Keitaro、Kawano Yoshiaki、Yamamoto Masaki、Hato Masakatsu、Nakamura Motonao、Shimizu Takao、Yokomizo Takehiko、Miyano Masashi、Yokoyama Shigeyuki
  • Journal Title: Nat Chem Biol Volume: 14 Pages: 262-269
  • DOI: 10.1038/nchembio.2547
  • Peer Reviewed / Open Access
• [Journal Article] Endurance exercise training and high-fat diet differentially affect composition of diacylglycerol molecular species in rat skeletal muscle. (2018)
  • Author(s): Kawanishi Noriaki、Takagi Kana、Lee Hyeon-Cheol、Nakano Daiki、Okuno Toshiaki、Yokomizo Takehiko、Machida Shuichi
  • Journal Title: Am J Physiol Regul Integr Comp Physiol Volume: 印刷中
  • DOI: 10.1152/ajpregu.00371.2017
  • Peer Reviewed / Open Access
• [Journal Article] Profiling of bioactive lipids in different dendritic cell subsets using an improved multiplex quantitative LC-MS/MS method (2018)
  • Author(s): Ohba M., Saeki K., Koga T., Okuno T., Kobayashi Y., Yokomizo T.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 504 Pages: 562-568
  • DOI: 10.1016/j.bbrc.2018.06.026
  • Peer Reviewed / Open Access
• [Journal Article] Altered eicosanoid production and phospholipid remodeling during cell culture. (2018)
  • Author(s): Okuno Toshiaki、Gij?n Miguel A.、Zarini Simona、Martin Sarah A.、Barkley Robert M.、Johnson Christopher A.、Ohba Mai、Yokomizo Takehiko、Murphy Robert C.
  • Journal Title: J Lipid Res Volume: 59 Pages: 542-549
  • DOI: 10.1194/jlr.m083030
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Leukotriene B4 promotes neovascularization and macrophage recruitment in murine wet-type AMD models (2018)
  • Author(s): Sasaki F., Koga T., Ohba M., Saeki K., Okuno T., Ishikawa K., Nakama T., Nakao S., Yoshida S., Ishibashi T., Ahmadieh H., Kanavi M. R., Hafezi-Moghadam A., Penninger J. M., Sonoda K. H., Yokomizo T.
  • Journal Title: JCI Insight Volume: 3 Pages: 96902-96902
  • DOI: 10.1172/jci.insight.96902
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Urinary prostaglandin D2 metabolite excretion during the first six months of life was significantly lower in breast-fed than formula-fed infants. (2018)
  • Author(s): Shoji Hiromichi、Ikeda Naho、Kitamura Tomohiro、Suganuma Hiroki、Ohba Mai、Okuno Toshiaki、Yokomizo Takehiko、Shimizu Toshiaki
  • Journal Title: Acta Paediatr Volume: 107 Pages: 95-100
  • DOI: 10.1111/apa.14068
  • Peer Reviewed / Open Access
• [Journal Article] Biological functions of 12(S)-hydroxyheptadecatrienoic acid as a ligand of leukotriene B4 receptor 2 (2018)
  • Author(s): Okuno T., Yokomizo T.
  • Journal Title: Inflammation and Regeneration Volume: 38 Pages: 29-29
  • DOI: 10.1186/s41232-018-0087-4
  • Peer Reviewed / Open Access
• [Journal Article] Non-steroidal anti-inflammatory drug delays corneal wound healing by reducing production of 12-hydroxyheptadecatrienoic acid a ligand for leukotriene B4 receptor 2 (2017)
  • Author(s): Iwamoto Satoshi、Koga Tomoaki、Ohba Mai、Okuno Toshiaki、Koike Masato、Murakami Akira、Matsuda Akira、Yokomizo Takehiko
  • Journal Title: Sci Rep Volume: 7 Pages: 1-10
  • DOI: 10.1038/s41598-017-13122-8
  • Peer Reviewed / Open Access
• [Journal Article] Biochemical and immunological characterization of a novel monoclonal antibody against mouseleukotriene B4 receptor 1. (2017)
  • Author(s): Sasaki F, Koga T, Saeki K, Okuno T, Kazuno S, Fujimura T, Ohkawa Y, Yokomizo T
  • Journal Title: PLoS One. Volume: 12 Pages: 0185133-0185133
  • DOI: 10.1371/journal.pone.0185133
  • Peer Reviewed / Open Access
• [Journal Article] Determination of Double Bond Positions in Polyunsaturated Fatty Acids Using the Photochemical Patern?-B?chi Reaction with Acetone and Tandem Mass Spectrometry (2017)
  • Author(s): Murphy Robert C.、Okuno Toshiaki、Johnson Christopher A.、Barkley Robert M.
  • Journal Title: Analytical Chemistry Volume: 89 Pages: 8545-8553
  • DOI: 10.1021/acs.analchem.7b02375
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Generation and characterization of a human-mouse chimeric high-affinity antibody that detects the DYKDDDDK FLAG peptide. (2017)
  • Author(s): Ikeda K, Koga T, Sasaki F, Ueno A, Saeki K, Okuno T, Yokomizo T.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 486 Pages: 1077-1082
  • DOI: 10.1016/j.bbrc.2017.03.165
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] ntravenous anesthetic propofol binds to 5-lipoxygenase and attenuates leukotriene B4 production. (2017)
  • Author(s): Okuno, T., Koutsogiannaki, S., Ohba, M., Chamberlain, M., Bu, W., Lin, F. Y., Eckenhoff, R. G., Yokomizo, T.,Yuki, K.
  • Journal Title: The FASEB Journal Volume: 31 Pages: 1584-1594
  • DOI: 10.1096/fj.201601095r
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Leukotriene B4 receptor type 2 protects against pneumolysin-dependent acute lung injury. (2017)
  • Author(s): Shigematsu, M., Koga, T., Ishimori, A., Saeki, K., Ishii, Y., Taketomi, Y., Ohba, M., Jo-Watanabe, A., Okuno, T., Harada, N., Harayama, T., Shindou, H., Li, J. D., Murakami, M., Hoka, S.,Yokomizo, T.
  • Journal Title: Scientific Reports Volume: 6 Pages: 34560-34560
  • DOI: 10.1038/srep34560
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Prolonged exposure to volatile anesthetic isoflurane worsens the outcome of polymicrobial abdominal sepsis. (2017)
  • Author(s): Koutsogiannaki, S., Schaefers, M. M., Okuno, T., Ohba, M., Yokomizo, T., Priebe, G. P., DiNardo, J. A., Sulpicio, S. G.,Yuki, K.
  • Journal Title: Toxicol Sci Volume: in press Pages: 402-411
  • DOI: 10.1093/toxsci/kfw261
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] chiki T, Koga T, Okuno T, Saeki K, Yamamoto Y, Yamamoto H, Sakaguchi M, Yokomizo T. Modulation of leukotriene B4 receptor 1 signaling by receptor for advanced glycation end products (RAGE). (2016)
  • Author(s): Ichiki T, Koga T, Okuno T, Saeki K, Yamamoto Y, Yamamoto H, Sakaguchi M, Yokomizo T
  • Journal Title: FASEB Journal Volume: 30 Pages: 1811-1822
  • DOI: 10.1096/fj.201500117
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Leukotriene B4 receptor type 2 (BLT2) enhances skin barrier function by regulating tight junction proteins. F (2016)
  • Author(s): Ishii Y, Saeki K, Liu M, Sasaki F, Koga T, Kitajima K, Meno C, Okuno T, Yokomizo T
  • Journal Title: Faseb J Volume: 30 Pages: 933-47
  • DOI: 10.1096/fj.15-279653
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] A fatty acid 12-hydroxyheptadecatrienoic acid accelerates corneal wound healing through leukotriene B4 receptor 2, BLT2. (2018)
  • Author(s): Yokomizo T., Iwamoto S., Koga T., Ohba M., Okuno T., Koike M., Murakami A., Matsuda A.
  • Organizer: Keystone Symposium, GPCR Structure and Function
  • Int'l Joint Research
• [Presentation] Dietary omega-3 fatty acids alleviate allergic conjunctivitis in a mouse model. (2018)
  • Author(s): Hirakata T., Lee C. H., Ohba M., Okuno T., Murakami A., Matsuda A., Yokomizo T.
  • Organizer: ARVO 2018
  • Int'l Joint Research
• [Presentation] Crystal structure of leukotriene B4 receptor 1 bound with an inverse agonist. (2018)
  • Author(s): Okuno T., Hori T., Yokoyama S., Yokomizo T.
  • Organizer: International GPCR symposium
  • Int'l Joint Research
• [Presentation] Phospholipid remodeling altered eicosanoid production during cell culture. (2018)
  • Author(s): Okuno T., Gijon M.A., Yokomizo T., Murphy R.C.
  • Organizer: ELM2018
  • Int'l Joint Research
• [Presentation] An algal precursor rearranges in a new 1,8-diol forming reaction to bioactive LTB4 enantiomers (2018)
  • Author(s): Jagusch H., Werner M., Okuno T., Yokomizo T., Werz O., Pohnert G.
  • Organizer: ELM2018
  • Int'l Joint Research
• [Presentation] アレルギー性結膜炎におけるオメガ3脂肪酸の効果の検証. (2018)
  • Author(s): 平形寿彬, 李賢哲, 大塲麻衣, 佐伯和子, 奥野利明, 松田彰, 村上晶, 横溝岳彦.
  • Organizer: 第60回日本脂質生化学会
• [Presentation] ロイコトリエンA4水解酵素の代謝における解析. (2018)
  • Author(s): 鵜澤博嗣, 古賀友紹, 奥野利明, 佐伯和子, 横溝岳彦.
  • Organizer: 第60回日本脂質生化学会
• [Presentation] 生理活性脂質12-HHTのBLT2受容体を介した生体保護的役割. (2018)
  • Author(s): 佐伯和子, 重松美沙子, 古賀友紹, 奥野利明, 横溝岳彦.
  • Organizer: 第91回日本生化学会
• [Presentation] Protective roles of 12-hydroxyheptadecatrienoic acid/BLT2 in acute lung injury. (2017)
  • Author(s): Shigematsu M, Koga T, Saeki K, Ohba M, Okuno T, Yokomizo T.
  • Organizer: Keystone Symposium, Lipidomics and Bioactive Lipids in Metabolism and Disease
  • Place of Presentation: Granlibakken Tahoe
  • Year and Date: 2017-02-26
  • Int'l Joint Research
• [Presentation] 静脈麻酔薬プロポフォールは5リポキシゲナーゼに直接結合しロイコトリエンB4産生を抑制する (2017)
  • Author(s): 奥野利明, 大塲麻生, Chamberlain M., Bu W., Liu F.-Y., Eckenhoff R. G., 横溝岳彦, 結城公一
  • Organizer: 第59回日本脂質生化学会
• [Presentation] 長期間の高強度持久的運動トレーニングによる骨格筋内のアシルカルニチン分子種濃度の変動 (2017)
  • Author(s): 川西範明, 高木香奈, 李賢哲, 奥野利明, 横溝岳彦, 町田修一
  • Organizer: 第72回日本体力医学会大会
• [Presentation] 麻酔薬プロポフォールの抗炎症作用機序の解明 (2017)
  • Author(s): 奥野利明,結城公一,横溝岳彦
  • Organizer: 第14回東京呼吸器リサーチフォーラム
• [Presentation] 静脈麻酔薬プロポフォールは5-リポキシゲナーゼに結合してロイコトリエンの産生を抑制する (2017)
  • Author(s): 奥野利明, Koutsogiannaki S., 大塲麻生, Chamberlain M., Bu W., Lin F.-Y., Eckenhoff R., 横溝岳彦, 結城公一
  • Organizer: 2017年度生命科学系学会合同年次大会(ConBio2017)
• [Presentation] Protective roles of 12-hydroxyheptadecatrienoic acid/BLT2 in acute lung injury (2017)
  • Author(s): Shigematsu M., Koga T., Saeki K., Ohba M., Okuno T., Yokomizo T
  • Organizer: Keystone Symposium, Lipidomics and Bioactive Lipids in Metabolism and Disease
  • Int'l Joint Research
• [Presentation] 12-HHT/BLT2 axis protects against pneumolysin-dependent acute lung injury (2017)
  • Author(s): Saeki K, Shigematsu M, Koga T, Ohba M, Okuno T, Yokomizo T
  • Organizer: Keystone Symposia Conference, Integrating Metabolism and Immunity
  • Int'l Joint Research
• [Presentation] Intravenous anesthetic propofol binds to 5-lipoxygenase and attenuates LTB4 production in vivo and in vitro. (2017)
  • Author(s): Okuno T., Koutsogiannaki S., Ohba M., Chamberlain M., Bu W., Lin F.-Y., Eckenhoff R. G., Yokomizo T., Yuki K
  • Organizer: 15th International Conference on Bioactive Lipids in Cancer, Inflammation and Related Diseases
  • Int'l Joint Research
• [Presentation] Dietary omega-3 fatty acids alleviate the allergic conjunctivitis in mice. (2017)
  • Author(s): Hirakata T., Lee C. H., Ohba M., Okuno T., Matsuda A., Murakami A., Yokomizo T.
  • Organizer: 15th International Conference on Bioactive Lipids in Cancer, Inflammation and Related Diseases
  • Int'l Joint Research
• [Presentation] RAGEはロイコトリエンB4第一受容体と機能的に相互作用する (2016)
  • Author(s): 市木貴子, 古賀友紹, 奥野利明, 佐伯和子, 阪口政清, 山本靖彦, 横溝岳彦.
  • Organizer: 第89回日本生化学会
  • Place of Presentation: 仙台
  • Year and Date: 2016-09-25
• [Presentation] The roles of 12-hydroxyheptadecatrienoic acid/BLT2 axis in skin. (2016)
  • Author(s): Shigematsu M, Koga T, Saeki K, Okuno T, Yokomizo T.
  • Organizer: 57th ICBL, International Conference on the Bioscience of Lipids
  • Place of Presentation: Chamonix-Mont-Blanc
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] 2-hydroxyheptadecatrienoic acid (12-HHT) as a novel lipid mediator. (2016)
  • Author(s): Yokomizo T, Y I, Shimegatsu M, Saeki K, Okuno T.
  • Organizer: 第58回日本脂質生化学会
  • Place of Presentation: 秋田
  • Year and Date: 2016-06-09
• [Presentation] 持久的運動トレーニングおよび高脂肪食餌負荷が骨格筋のリン脂質濃度に及ぼす影響 (2016)
  • Author(s): 川西範明, 高木香奈, 李賢哲, 奥野利明, 横溝岳彦, 町田修一
  • Organizer: 第58回日本脂質生化学会
  • Place of Presentation: 秋田
  • Year and Date: 2016-06-09
• [Presentation] Receptor for advanced glycation endproducts modulates leukotriene B4 receptor 1 signaling. (2016)
  • Author(s): Ichiki T, Koga T, Okuno T, Saeki K, Yamamoto Y, Sakaguchi M, Yokomizo T.
  • Organizer: ASBMB meeting
  • Place of Presentation: San Diego
  • Year and Date: 2016-04-02
  • Int'l Joint Research
• [Presentation] Generation of chimeric high-affinity monoclonal antibody for FLAG tag. (2016)
  • Author(s): Ikeda K, Sasaki F, Ueno A, Okuno T, Koga T, Yokomizo T.
  • Organizer: ASBMB meeting
  • Place of Presentation: San Diego
  • Year and Date: 2016-04-02
  • Int'l Joint Research
• [Presentation] BLT2 receptor protects against lethal acute lung injury induced by a pneumococcal toxin pneumolysin. (2016)
  • Author(s): Shigematsu M, Koga T, Saeki K, Okuno T, Yokomizo T.
  • Organizer: ASBMB meeting
  • Place of Presentation: San Diego
  • Year and Date: 2016-04-02
  • Int'l Joint Research
• [Remarks] 順天堂大学医学部生化学第一講座 Publication List
  • URL: http://plaza.umin.ac.jp/j_bio/publication.html
• [Remarks] Publication List
  • URL: http://plaza.umin.ac.jp/j_bio/publication.html
• [Remarks] researchmap 奥野利明
  • URL: http://researchmap.jp/toshiakiokuno