Elucidation of the correlation between the structure of the membrane protein LMP1 derived from EB virus and its function of latent infection

• Project/Area Number: 25282230
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: Osaka University
• Principal Investigator: Inoue Tsuyoshi 大阪大学, 工学(系)研究科(研究院), 教授 (20263204)
• Co-Investigator(Kenkyū-buntansha): 安居 輝人 大阪大学, 微生物病研究所, 准教授 (60283074)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2016: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2015: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2014: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: Epstein-Barr（EB）ウイルス / 潜伏感染 / 不死化機構 / LMP1 / LMP2A / 膜蛋白質 / 自己免疫疾患 / リンパ腫 / Ⅹ線構造解析 / 精製 / 結晶化 / EBウィルス / 膜タンパク質 / X線構造解析 / 抗体 / ヘルペスウイルス / H-PGDS

Outline of Final Research Achievements

Mice expressing membrane proteins LMP1 and LMP2A derived from EV virus were prepared and their effects were investigated. It was found that LMP2A promotes the differentiation of germinal center B cells into antibody producing cells and contributes to infection of EBV by surviving low affinity B cells. In addition, systemic lupus erythematosus-like symptoms were induced and firstly proved to be associated with autoimmune diseases.In addition, T / NK cells were reduced by using its antibody, and the pathological condition of the mouse was analyzed in the state of immunosuppression. It was found that LMP1 and LMP2A cooperate with each other in germinal center B cells and induce proliferative diseases such as Hodgkin's lymphoma.
The crystallization of the C-terminus region of CTAR1 involved in signal transduction in LMP1 was performed. The complex crystals between CTAR1 and its antibody of 11A1 up to 1.9 A resolution were obtained and the structural analysis is now in progress.

Research Products

• [Journal Article] A RuBisCO-mediated carbon metabolic pathway in methanogenic archaea (2017)
  • Author(s): T. Kono, S. Mehrotra, C. Endo, N. Kizu, M. Matsuda, H. Kimura, E. Mizohata, T. Inoue, T. Hasunuma, A. Yokota, H. Matsumura, H. Ashida
  • Journal Title: Nature Commun. Volume: 8 Issue: 1 Pages: 14007-14007
  • DOI: 10.1038/ncomms14007
  • NAID: 120005947453
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] A trimeric structural fusion of an antagonistic tumor necrosis factor-α mutant enhances molecular stability and enables facile modification (2017)
  • Author(s): Masaki Inoue, Daisuke Ando, Haruhiko Kamada, Shintaro Taki, Mayumi Niiyama, Yohei Mukai, Takashi Tadokoro, Katsumi Maenaka, Taisuke Nakayama, Yuji Kado, Tsuyoshi Inoue, Yasuo Tsutsumi and Shin-ichi Tsunoda
  • Journal Title: J. Biol. Chem., Volume: in press Issue: 16 Pages: 235-237
  • DOI: 10.1074/jbc.m117.779686
  • Peer Reviewed / Open Access
• [Journal Article] New molecular packing in a crystal of pseudoazurin from Alcaligenes faecalis: a double-helical arrangement of blue copper. (2017)
  • Author(s): Yohta Fukuda, Eiichi Mizohata and Tsuyoshi Inoue
  • Journal Title: Acta Cryst. F Volume: 73 Issue: 3 Pages: 159-166
  • DOI: 10.1107/s2053230x17002631
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Protein crystallization with paper (2016)
  • Author(s): Miki Matsuoka, Keisuke Kakinouchi, Hiroaki Adachi, Mihoko Maruyama, Shigeru Sugiyama, Satoshi Sano, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Masashi Yoshimura, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori and Kazufumi Takano
  • Journal Title: Jpn. J. Appl. Phys., Volume: 55 Issue: 5 Pages: 050302-050302
  • DOI: 10.7567/jjap.55.050302
  • NAID: 210000146451
  • Peer Reviewed / Open Access
• [Journal Article] Redox-coupled structural changes in nitrite reductase revealed by serial femtosecond and microfocus crystallography. (2016)
  • Author(s): Fukuda Y, Tse KM, Suzuki M, Diederichs K, Hirata K, Nakane T, Sugahara M, Nango E, Tono K, Joti Y, Kameshima T, Song C, Hatsui T, Yabashi M, Nureki O, Matsumura H, Inoue T, Iwata S, Mizohata E.
  • Journal Title: J. Biochem Volume: 未定 Issue: 5 Pages: 527-538
  • DOI: 10.1093/jb/mvv133
  • NAID: 40020833586
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular mechanism underlying promiscuous polyamine recognition by spermidine acetyltransferase (2016)
  • Author(s): S. Sugiyama, S. Ishikawa, H. Tomitori, M. Niiyama, M. Hirose, Y. Miyazaki, K. Higashi, M. Murata, H. Adachi, K. Takano, S. Murakami, T. Inoue, Y. Mori, K. Kashiwagi, K. Igarashi, H. Matsumura
  • Journal Title: Int. J. Biochem. Cell Biol. Volume: 76 Pages: 87-97
  • DOI: 10.1016/j.biocel.2016.05.003
  • Peer Reviewed / Open Access
• [Journal Article] A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique (2016)
  • Author(s): M. Maruyama, Y. Hayashi, H. Y. Yoshikawa, S. Okada, H. Koizumi, M. Tachibana, S. Sugiyama, H. Adachi, H. Matsumura, T. Inoue, K. Takano, S. Murakami, M. Yoshimura, Y. Mori
  • Journal Title: J. Crystal Growth Volume: 452 Pages: 172-178
  • DOI: 10.1016/j.jcrysgro.2015.11.008
  • Peer Reviewed / Open Access
• [Journal Article] Promotion of protein crystal growth by actively switching crystal growth mode via femtosecond laser ablation (2016)
  • Author(s): Y. Tominaga, M. Maruyama, M. Yoshimura, H. Koizumi, M. Tachibana, S. Sugiyama, H. Adachi, K. Tsukamoto, H. Matsumura, K. Takano, S. Murakami, T. Inoue, H. Y. Yoshikawa, Y. Mori
  • Journal Title: Nature Photonics Volume: 10 Issue: 11 Pages: 723-726
  • DOI: 10.1038/nphoton.2016.202
  • Peer Reviewed / Open Access
• [Journal Article] Ubiquitination of Lysine 867 of the Human SETDB1 Protein Upregulates Its Histone H3 Lysine 9 (H3K9) Methyltransferase Activity. (2016)
  • Author(s): Ishimoto K, Kawamata N, Uchihara Y, Okubo M, Fujimoto R, Gotoh E, Kakinouchi K, Mizohata E, Hino N, Okada Y, Mochizuki Y, Tanaka T, Hamakubo T, Sakai J, Kodama T, Inoue T, Tachibana K, Doi T
  • Journal Title: PLoS One Volume: 11 Issue: 10 Pages: e0165766-e0165766
  • DOI: 10.1371/journal.pone.0165766
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Epiregulin Recognition Mechanisms by Anti-epiregulin Antibody 9E5: STRUCTURAL, FUNCTIONAL, AND MOLECULAR DYNAMICS SIMULATION ANALYSES. (2016)
  • Author(s): Kado Y1, Mizohata E2, Nagatoishi S3, Iijima M4, Shinoda K4, Miyafusa T3, Nakayama T2, Yoshizumi T2, Sugiyama A4, Kawamura T4, Lee YH4, Matsumura H2, Doi H4, Fujitani H4, Kodama T4, Shibasaki Y4, Tsumoto K5, Inoue T6.
  • Journal Title: J Biol Chem. Volume: 291 Issue: 5 Pages: 2319-2330
  • DOI: 10.1074/jbc.m115.656009
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure-based design and synthesis of a bivalent iminobiotin analog showing strong affinity toward a low immunogenic streptavidin mutant. (2015)
  • Author(s): Kawato T, Mizohata E, Shimizu Y, Meshizuka T, Yamamoto T, Takasu N, Matsuoka M, Matsumura H, Kodama T, Kanai M, Doi H, Inoue T, Sugiyama A
  • Journal Title: Biosci Biotechnol Biochem. Volume: 79 Issue: 4 Pages: 640-642
  • DOI: 10.1080/09168451.2014.991692
  • Peer Reviewed / Open Access
• [Journal Article] The N-terminal acidic residue of the cytosolic helix 8 of an odorant receptor is responsible for different response dynamics via G-protein (2015)
  • Author(s): T. Kawasaki, T. Saka, S. Mine, E. Mizohata, T. Inoue, H. Matsumura, T. Sato
  • Journal Title: FEBS Lett. Volume: 589 Issue: 10 Pages: 1136-1143
  • DOI: 10.1016/j.febslet.2015.03.025
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Development of protein seed crystals reinforced by high-strength hydrogels (2015)
  • Author(s): Shigeru Sugiyama, Noriko Shimizu, Keisuke Kakinouchi, Osamu Hiraoka, Hiroyoshi Matsumura, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Mihoko Maruyama, Masashi Yoshimura, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Michio Murata, and Yusuke Mori
  • Journal Title: CrystEngComm Volume: 17 Issue: 42 Pages: 8064-8071
  • DOI: 10.1039/c5ce00844a
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Insights into unknown foreign ligand in copper nitrite reductase (2015)
  • Author(s): Yohta Fukuda, Tse Ka Man, Yuji Kado, Eiichi Mizohata, Hiroyoshi Matsumura, Tsuyoshi Inoue
  • Journal Title: Biochem. Biophys. Res. Commun., Volume: 464（２） Issue: 2 Pages: 622-628
  • DOI: 10.1016/j.bbrc.2015.07.025
  • Peer Reviewed / Open Access
• [Journal Article] Redox-coupled proton transfer mechanism in nitrite reductase revealed by femtosecond crystallography (2015)
  • Author(s): Yohta Fukuda, Ka Man Tse, Takanori Nakane, Toru Nakatsu, Mamoru Suzuki, Michihiro Sugahara, Shigeyuki Inoue, Tetsuya Masuda, Fumiaki Yumoto, Naohiro Matsugaki, Eriko Nango, Tsuyoshi Inoue, So Iwata, Eiichi Mizohata、et. el
  • Journal Title: PNAS., Volume: 55 Pages: 050302-050302
  • Peer Reviewed / Open Access
• [Journal Article] Laser ablation for protein crystal nucleation and seeding (2014)
  • Author(s): H.Y.Yoshikawa, R.Murai, H.Adachi, S.Sugiyama, M.Maruyama, Y.Takahashi, K.Takano, H.Matsumura, T.Inoue, S.Murakami, H.Masuhara, and Y.Mori
  • Journal Title: Chemical Society Reviews Volume: Vol.43, No.7 Issue: 7 Pages: 2147-2158
  • DOI: 10.1039/c3cs60226e
  • Peer Reviewed
• [Journal Article] A new practical technique for high quality protein crystallization with the solution stirring technique at the interface between high-concentrated hydrogel and solution (2014)
  • Author(s): Y. Aoki, M. Maruyama, Y. Takahashi, M. Yoshimura, H. Y. Yoshikawa, S. Sugiyama, H. Adachi, K. Takano, S. Murakami, H. Matsumura, T. Inoue and Y. Mori,
  • Journal Title: Jpn. J. Appl. Phys Volume: (in press)
  • NAID: 210000144031
  • Peer Reviewed
• [Journal Article] A novel approach for protein crystallization by a synthetic hydrogel with thermoreversible gelation polymer (2013)
  • Author(s): S. Sugiyama, N. Shimizu, G. Sazaki, M. Hirose, Y. Takahashi, M. Maruyama, H. Matsumura, H. Adachi, K. Takano, S. Murakami, T. Inoue, and Y. Mori,
  • Journal Title: Cryst. Growth Des. Volume: 13 Issue: 5 Pages: 1899-1904
  • DOI: 10.1021/cg301588b
  • Peer Reviewed
• [Presentation] Structural Bioinorganic Chemistry of Copper Enzymes (2016)
  • Author(s): Yohta Fukuda, Eiichi Mizohata, Tsuyoshi Inoue
  • Organizer: The 4th International Symposium for Young Chemists on Stimuli-Responsive Chemical Species for the Creation of Functional Molecules
  • Place of Presentation: 大阪大学吹田キャンパス 工学研究科サントリーメモリアルホール（大阪府・吹田市）
  • Year and Date: 2016-12-12
  • Int'l Joint Research / Invited
• [Presentation] 造血器型プロスタグランジンD合成酵素変異体V187Iの熱安定性 (2016)
  • Author(s): 門 祐示、Pojchanun Kanitthamniyom、 中村 努、 Sun Xin、 中山 泰亮、 福田 庸太、 溝端 栄一、裏出 良博、 井上 豪
  • Organizer: 日本結晶学会平成28年度年会
  • Place of Presentation: 茨城県立県民文化センター（茨城県・水戸市）
  • Year and Date: 2016-11-17
• [Presentation] New Hot Topics on Copper Nitrite Reductases (2016)
  • Author(s): Yohta Fukuda
  • Organizer: 5th International Symposium on Diffraction Structural Biology (ISDSB2016)
  • Place of Presentation: Hilton Downtown Knoxville, TN, US（アメリカ合衆国・テネシー州）
  • Year and Date: 2016-08-07
  • Int'l Joint Research / Invited
• [Presentation] SACLA が解明した銅含有亜硝酸還元酵素の常温無損傷構造 (2016)
  • Author(s): 溝端 栄一
  • Organizer: 第16回日本蛋白質科学会年会
  • Place of Presentation: 福岡国際会議場（福岡県・福岡市）
  • Year and Date: 2016-06-07