ＳＦＸを用いたＨＩＶアクセサリータンパク質と宿主制限因子の複合体の動的構造の解明

• Project/Area Number: 16F16111
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Physical pharmacy
• Research Institution: Kumamoto University
• Principal Investigator: 大塚 雅巳 熊本大学, 大学院生命科学研究部(薬), 教授 (40126008)
• Co-Investigator(Kenkyū-buntansha): CIFTCI HALIL 熊本大学, 大学院生命科学研究部(薬), 外国人特別研究員
• Project Period (FY): 2016-04-22 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥2,300,000 (Direct Cost: ¥2,300,000); Fiscal Year 2017: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2016: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: HIV / MA / IP6 / SFX / Vpx

Outline of Annual Research Achievements

①SAMHD1およびDCAF1タンパク質を発現し精製することに成功したが、アクセサリータンパク質Vpxを精製するには至らなかった。E. coliの系統を変える、GSTタグをもつVpxタンパク質を構築するなど試みたが、結晶精製スクリーニングに必要なVpx-SAMHD1-DCAF1三者複合体を形成するに十分な濃度のVpxタンパク質を得るに至らなかった。
②MA-IP6複合体結晶を得ることができ、2つの異なった形の結晶を回折実験に供し、2.7および2.5 Åの分解能で結晶構造を解くことができた。予想したとおり、IP6分子はタンパク質の塩基性の高い領域のアミノ酸残基と相互作用していた。両結晶のパッキングについてはMAの集合とIP6分子との観点から議論することができる。現在、スタンフォード大学との共同研究により、大阪のSACLAの2018年5月のビームタイムにおいてIP6のナノ結晶の室温におけるSFX実験を行っているところである。
③トルコ薬用植物と合成化合物を基軸とした医薬スクリーニングについては、benzyl 3-hydroxy-23-oxoolean-12-en-28-oateがBcr-Abl陽性のK562細胞に対して最も有効で（IC50は9.3 μM）、腫瘍特異性とアポプトーシス誘導活性を示した。本化合物はAblプロテインキーゼを低いマイクロモルオーダーのIC50値で阻害するが、BRK, BTK, CSK, LYN BおよびSRCキナーゼに対する阻害効果はイマチニブより高いものであった。さらに、本化合物とBcr-Ablの複合体のドッキングシミュレーションの結果ベンジルエステル部分が幾つかの重要なアミノ酸残基と結合および疎水性相互作用を持つことが示された。全体として、本研究でジプソジェニン誘導体がキナーゼ阻害剤の新しいクラスとなることを示した。

Research Progress Status

29年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

29年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Anadol University/Ege University(Turkey)
• [Int'l Joint Research] Stanford University(米国)
• [Journal Article] The First Pentacyclic Triterpenoid Gypsogenin Derivative Exhibiting Anti-ABL1 Kinase and Anti-chronic Myelogenous Leukemia Activities (2018)
  • Author(s): Ciftci, H.I., Ozturk, S.E., Ali, T.F.S., Radwan, M.O., Tateishi, H., Koga, R., Ocak, Z., Can, M., Otsuka, M., and Fujita, M.
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 41 Issue: 4 Pages: 570-574
  • DOI: 10.1248/bpb.b17-00902
  • NAID: 130006602535
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Introduction of H2C2-type zinc-binding residues into HIV-2 Vpr increases its expression level. (2018)
  • Author(s): Koga, R., Yamamoto, M., Ciftci, H.I., Otsuka, M., and Fujita, M.
  • Journal Title: FEBS Open Bio Volume: 1 Issue: 1 Pages: 146-153
  • DOI: 10.1002/2211-5463.12358
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Design, synthesis and biological evaluation of novel 1,3,4-Thiadiazole derivatives as potential antitumor agents against chronic myelogenous leukemia. Striking effect of nitrothiazole moiety. (2018)
  • Author(s): Altintop, M.D., Ciftci, H.I., Radwan, M.O., Sever, B., Kaplancikli, Z.A., Ali, T.F.S., Koga, R., Fujita, M., Otsuka, M., and Ozdemir, A.
  • Journal Title: Molecules Volume: 23 Issue: 1 Pages: 59-59
  • DOI: 10.3390/molecules23010059
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A dithiol compound binds to the zinc finger protein TRAF6 and suppresses its ubiquitination. (2017)
  • Author(s): Koga, R., Radwan, M.O., Ejima, T., Kanemaru, Y., Tateishi, H., Ali, T.F.S., Ciftci, H.I., Shibata, Y., Taguchi, Y., Inoue, J., Otsuka, M., and Fujita, M.
  • Journal Title: ChemMedChem Volume: 12 Issue: 23 Pages: 1935-1941
  • DOI: 10.1002/cmdc.201700399
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A clue to unprecedented strategy to HIV eradication: “Lock-in and apoptosis”. (2017)
  • Author(s): Tateishi, H., Monde, K., Anraku, K., Koga, R., Hayashi, Y., Ciftci, H.I., DeMirci, H., Higashi, T., Motoyama, K, Arima, H., Otsuka, M., and Fujita, M.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 8957-8957
  • DOI: 10.1038/s41598-017-09129-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Synthesis, Computational Study, and Evaluation of in vitro Antimicrobial, Antibiofilm, and Anticancer Activities of New Sulfanyl Aminonaphthoquinone Derivatives (2017)
  • Author(s): Nilufer Bayrak, Hatice Yildirim, Amac Fatih Tuyun, Emel Mataraci Kara, Berna Ozbek Celik, Girish Kumar Gupta, Halil I. Ciftci, Mikako Fujita, Masami Otsuka, Hamid R. Nasiri
  • Journal Title: Letters in Drug Design & Discovery, Volume: 14 Issue: 6 Pages: 647-661
  • DOI: 10.2174/157018081406170606155530
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Se-SAD serial femtosecond crystallography dataset from selenobiotinyl-streptavidin (2017)
  • Author(s): Chun Hong Yoon, Hasan DeMirci, Raymond G. Sierra, E. Han Dao, Radman Admadi, Fulya Aksit, Andrew L. Aquila, Alexander Batyuk, Halil I Ciftci, et al.
  • Journal Title: Scientific Data Volume: 4 Issue: 1 Pages: 170055-170055
  • DOI: 10.1038/sdata.2017.55
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Role of zinc binding site of HIV-2 Vpx prevents instability and dysfunction of the protein (2017)
  • Author(s): Minami Yamamoto, Ryoko Koga, Haruna Fujino, Kazunori Shimagaki, Halil Ibrahim Ciftci, Masahiro Kamo, Hiroshi Tateishi, Masami Otsuka, Mikako Fujita
  • Journal Title: J. General Virol. Volume: 98 Issue: 2 Pages: 275-283
  • DOI: 10.1099/jgv.0.000701
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Zinc binding site of HIV-2 Vpx prevents instability and dysfunction of the protein. (2017)
  • Author(s): 3.Yamamoto M, Koga R, Fujino H, Shimagaki K, Ciftci H.I, Kamo M, Tateishi H, Otsuka M and Fujita M.
  • Journal Title: Journal of General Virology Volume: 98 Pages: 275-283
  • Peer Reviewed
• [Journal Article] Se-SAD serial femtosecond crystallography datasets from selenobiotinyl-streptavidin. (2017)
  • Author(s): 4.Chun HY, Hasan D, Raymond GS, E Han D, Radman A, Fulya A, Andrew LA, Alexander B, Halilibrahim C, Serge G, Matt JH, Brandon H, Thomas JL, Meng L, Ulf L, Jason EK, Paul M, Yashas R, Theodore R, Evan R, Lindsey Z, Soichi W, Sebastien B, James MH, Mark SH.
  • Journal Title: Scientific Data Volume: 4 Pages: 170055-170055
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Selenium single-wavelength anomalous diffraction de novo phasing using an X-ray Free Electron Laser. (2016)
  • Author(s): 1.Mark Hunter, Chun Yoon, Hasan DeMirci, Raymond Sierra, Eugene Dao, Radman Admadi, Fulya Aksit, Andrew Aquila, Halilibrahim Ciftci, Serge Guillet, Matt Hayes, Thomas Lane, Mengning Liang, Ulf Lundstrom, Jason Koglin, Paul Mgbam, Yashas Rao, Lindesy Zhang, Soichi Wakatsuki, James Holton, Sebastien Boutet.
  • Journal Title: Nature Communications Volume: 7 Pages: 13388-13388
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Kinase Inhibitory Activity of Glycyrrhetic Acid (2018)
  • Author(s): Halil I Ciftci, Mohamed O Radwan, Masami Otsuka, Mikako Fujita
  • Organizer: The 4th International Symposium on Pharmaceutical and Biomedical Sciences
  • Int'l Joint Research
• [Presentation] tructural Basis of Inositol Phosphate Recognition in the HIV-1 Gag Matrix (2017)
  • Author(s): Halil I Ciftci, Hiroshi Tateishi, Kotaro Koiwai, Ryoko Koga, Masami Otsuka, Mikako Fujita, Fumiaki Yumoto, Toshiya Senda
  • Organizer: Consortium of Biological Sciences
• [Presentation] Structural Basis of HIV-1 Gag Matrix Targeting by IP6 Derivative (2017)
  • Author(s): Halil I Ciftci, Hiroshi Tateishi, Kotaro Koiwai, Ryoko Koga, Masami Otsuka, Mikako Fujita, Fumiaki Yumoto, Toshiya Senda
  • Organizer: The 31st Annual Meeting of Japanese Society for AIDS Research
• [Presentation] Structural Basis of the Interaction between HIV-1 Gag Matrix and IP6 -Toward the Development of IP6 Derivative- (2017)
  • Author(s): Halil I Ciftci, Hiroshi Tateishi, Kotaro Koiwai, Ryoko Koga, Masami Otsuka, Mikako Fujita, Fumiaki Yumoto, and Toshiya Senda
  • Organizer: The 67th Annual Meeting of the Japanese Society for Virology
• [Presentation] Development of Anti-HIV and Anti-Cancer Drug Candidates (2017)
  • Author(s): Halil I Ciftci, Kotaro Koiwai, Hiroshi Tateishi, Mehlika D Altintop, Mohamed O Radwan, Belgin Sever, Taha FS Ali, Ryoko Koga, Zafer A Kaplancikli, Ahmet Ozdemir, Fumiaki Yumoto, Toshiya Senda, Mikako Fujita, Masami Otsuka
  • Organizer: Turkish Japanese Workshop on Bioorganic Medicinal Chemistry and Natural Products
  • Int'l Joint Research
• [Presentation] Structural Basis of Targeting HIV-1 Gag Matrix. Toward Finding a cure for HIV Infection (2017)
  • Author(s): Halil I Ciftci, Hiroshi Tateishi, Kotaro Koiwai, Ryoko Koga, Fumiaki Yumoto, Toshiya Senda, Masami Otsuka, and Mikako Fujita
  • Organizer: 2nd Japan Turkey International Symposium on Pharmaceutical and Biomedical Sciences (JATUSPAB-2)
  • Int'l Joint Research
• [Presentation] Kinase Inhibitory Activity of Gypsogenin Derivatives (2017)
  • Author(s): Halil I Ciftci, Safiye E Ozdemir, Taha FS Ali, Mohamed O Radwan, Hiroshi Tateishi, Ryoko Koga, Mustafa Can, Masami Otsuka, and Mikako Fujita
  • Organizer: 2nd Japan Turkey International Symposium on Pharmaceutical and Biomedical Sciences (JATUSPAB-2)
  • Int'l Joint Research
• [Presentation] Role of zinc binding site of HIV-2 Vpx (2016)
  • Author(s): Minami Yamamoto, Ryoko Koga, Haruna Fujino, Kazunori Shimagaki, Halil Ibrahim Ciftci, Masahiro Kamo, Hiroshi Tateishi, Masami Otsuka, Mikako Fujita
  • Organizer: 2 nd Kumamoto IRCMS International Symposium and 17 th Kumamoto AIDS Seminar
  • Place of Presentation: 熊本県民交流館パレア
  • Year and Date: 2016-10-31
  • Int'l Joint Research
• [Presentation] Characters of largely overexpressed HIV accessory proteins Vpx and Vif (2016)
  • Author(s): Kazunori Shimagaki, Halil Ibrahim Ciftci, Ryoko Koga, Haruna Fujino, Minami Yamamoto, Yoshihiro Yamaguchi, Masami Otsuka, Mikako Fujita
  • Organizer: 第64回日本ウイルス学会学術集会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2016-10-23
• [Presentation] Significance of Zinc Binding Site of HIV-2 Vpx (2016)
  • Author(s): Minami Yamamoto, Ryoko Koga, Haruna Fujino, Kazunori Shimagaki, Halil Ibrahim Ciftci, Masahiro Kamo, Hiroshi Tateishi, Masami Otsuka, Mikako Fujita
  • Organizer: Japan-Turkey International Symposium on Pharmaceutical and Biomedical Sciences
  • Place of Presentation: 熊本大学薬学部宮本記念館
  • Year and Date: 2016-10-02
  • Int'l Joint Research
• [Presentation] N -Substituted-L-Histidine Analogues as Anticancer Agents: A Structure-Activity Relationship Study (2016)
  • Author(s): Taha F. S. Ali, Kana Iwamaru, Halil Ibrahim Ciftci, Ryoko Koga, Hiromasa Kurosaki, Mikako Fujita, Yoshinari Okamoto, Masami Otsuka
  • Organizer: Japan-Turkey International Symposium on Pharmaceutical and Biomedical Sciences
  • Place of Presentation: 熊本大学薬学部宮本記念館
  • Year and Date: 2016-10-02
  • Int'l Joint Research
• [Presentation] Restructuring of the bleomycin Metal Core (2016)
  • Author(s): Taha F. S. Ali, Satomi Ida, Yosuke Kanemaru, Ryoko Koga, Ayumi Tanaka, Akiyuki Hamasaki, Tomohiko Ejima, Mohamed Osman Radwan, Halil Ibrahim Ciftci, Hiromasa Kurosaki, Yoshinari Okamoto, Mikako Fujita, Masami Otsuka
  • Organizer: Japan-Turkey International Symposium on Pharmaceutical and Biomedical Sciences
  • Place of Presentation: 熊本大学薬学部宮本記念館
  • Year and Date: 2016-10-02
  • Int'l Joint Research
• [Presentation] Functional and Structural Study of HIV Accessory Proteins with Anti-Viral Factors and Screening of New Anti-Cancer Inhibitors (2016)
  • Author(s): Halil Ibrahim Ciftci
  • Organizer: Japan-Turkey International Symposium on Pharmaceutical and Biomedical Sciences
  • Place of Presentation: 熊本大学薬学部宮本記念館
  • Year and Date: 2016-10-02
  • Int'l Joint Research / Invited
• [Remarks] 熊本大学大学院薬学教育部 生体機能分子合成学分野
  • URL: http://www.pharm.kumamoto-u.ac.jp/Labs/bunsigouseiHP/index.html
• [Remarks] 熊本大学大学院生命科学研究部 生体機能分子合成学分野
  • URL: http://www.medphas.kumamoto-u.ac.jp/research/bunya/60.html
• [Remarks] 生体機能分子合成学分野
  • URL: http://www.pharm.kumamoto-u.ac.jp/Labs/bunsigouseiHP/index.html