Elucidation of the Function of Vaults based on its whole structure.

• Project/Area Number: 24687020
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Biophysics
• Research Institution: Osaka University
• Principal Investigator: TANAKA Hideaki 大阪大学, たんぱく質研究所, 准教授 (40346169)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥27,820,000 (Direct Cost: ¥21,400,000、Indirect Cost: ¥6,420,000); Fiscal Year 2014: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000); Fiscal Year 2012: ¥14,560,000 (Direct Cost: ¥11,200,000、Indirect Cost: ¥3,360,000)
• Keywords: ボルト / 脂質ラフト / 自然免疫 / X線結晶構造解析

Outline of Final Research Achievements

Vault is the largest cytoplasmic nucleoprotein particle. However we have determined the x-ray structure of rat liver vault at 3.5 angstrom resolution in 2009 (Science 323 384-288, 2009), the electron density map around the waist domain (N terminal domain of MVP) was disordered. Thus we added leucine zipper to the N-terminus of MVP (LZ-MVP), and expressed LZ-MVP in the Sf9 insect cell line using a baculovirus vector. We obtained more stable vault particles (LZ-vaults) in high purity and yield (80mg LZ-vaults/1L culture). The crystals of LZ vaults are diffracted up to 2.8angstrom resolution on beam line BL44XU at SPring-8.

Research Products

• [Journal Article] New features of vault architecture and dynamic srevealed by novel refinement using the deformable elastic network approach (2013)
  • Author(s): Casanas A, Querol-Audi J, Guerra P, Pous J, Tanaka H, Tsukihara T, Verdaguer N, Fita I
  • Journal Title: Acta Crystallogr D Biol Crystallogr Volume: 69 Issue: 6 Pages: 1054-1061
  • DOI: 10.1107/s0907444913004472
  • Peer Reviewed
• [Journal Article] Structural studies of large nucleoprotein particles, vaults. (2012)
  • Author(s): Tanaka H and Tsukihara T
  • Journal Title: Proc. Jpn. Acad., Ser. B Volume: 88 Pages: 416-433
  • NAID: 130001924758
  • Peer Reviewed
• [Presentation] Crystal Structure of Photosystem I from Synechocystis sp. PCC6803 at 5.1 Å Resolution (2015)
  • Author(s): 河合(久保田)寿子、坂本由佳、和田元、田中秀明、栗栖源嗣
  • Organizer: 日本化学会第95春季年会
  • Place of Presentation: 日本大学 理工学部船橋キャンパス／薬学部
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] 光化学系Iとフェレドキシンが形成する電子伝達複合体の構造基盤 (2014)
  • Author(s): 河合（久保田）寿子、武藤梨沙、池上貴久、Marc Nowaczyk、Matthias Rögner、田中秀明、栗栖源嗣
  • Organizer: 平成26年度日本結晶学会年会
  • Place of Presentation: 東京大学農学部
  • Year and Date: 2014-11-01 – 2014-11-03
• [Presentation] X-ray structure and NMR analysis of the electron transfer complex between photosystem I and ferredoxin (2014)
  • Author(s): Risa Mutoh, Hisako Kubota-Kawai, Marc Nowaczyk, Matthias Rögner, Hideaki Tanaka, Takahisa Ikegami, Genji Kurisu
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] 光化学系I－フェレドキシン複合体の構造解析 (2014)
  • Author(s): 武藤梨沙、久保田（河合）寿子、田中秀明、池上貴久、栗栖源嗣
  • Organizer: 第22回 「光合成セミナー2014：反応中心と色素系の多様性」
  • Place of Presentation: 名古屋工業大学
  • Year and Date: 2014-07-12 – 2014-07-13
• [Presentation] ロイシンジッパー付加によるボルト粒子の安定化と発現量の向上 (2013)
  • Author(s): 田中秀明、藤田千鶴子、岩崎憲治、月原冨武
  • Organizer: 第13回日本蛋白質科学会年会
  • Place of Presentation: とりぎん文化会館（鳥取県鳥取市）
• [Presentation] 生体粒子ボルトを利用したナノカプセルの開発 (2012)
  • Author(s): 田中秀明
  • Organizer: 第1回ネイチャーインダストリーアワード-若手研究者からの発信-
  • Place of Presentation: 大阪科学技術センター(大阪府大阪市)
  • Year and Date: 2012-11-20
• [Patent(Industrial Property Rights)] Artificial Bioparticle and Method of Manufacturing the Same (2015)
  • Inventor(s): 田中秀明
  • Industrial Property Rights Holder: 田中秀明
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-03-20
  • Overseas
• [Patent(Industrial Property Rights)] 人工生体粒子およびその製造方法 (2012)
  • Inventor(s): 田中秀明
  • Industrial Property Rights Holder: JST
  • Industrial Property Number: 2012-253031
  • Filing Date: 2012-11-19