Structural analysis of pore forming CEL-III complex.

• Project/Area Number: 20770088
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: Nagasaki University
• Principal Investigator: UNNO Hideaki Nagasaki University, 工学部, 助教 (10452872)
• Project Period (FY): 2008 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2009: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2008: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: CEL-III / 結晶構造解析 / PFT / Cucumaria echinata / 膜蛋白質

Research Abstract

For structural analysis of pore forming CEL-III complex, we tried to crystallize the complex and succeeded to make it plate-like crystal. Diffraction data from the complex crystal were collected by using synchrotron radiation in which the highest resolution leached to seven angstroms. Further trial of the crystallization using several kinds of detergent lead to obtain new crystals.

Research Products

• [Journal Article] The role and structure of the carboxyl-terminal domain of the human voltage-gated proton channel Hv1. (2010)
  • Author(s): Shu Jie Li, Qing Zhao, Qiangjun Zhou, Hideaki Unno, Yujia Zhai, Fei Sun
  • Journal Title: J. Biol. Chem. (In press)
  • Peer Reviewed
• [Journal Article] Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation (2010)
  • Author(s): Norihisa Yasui, Emiko Mihara, Maiko Nampo, Keiko Tamura-Kawakami, Hideaki Unno, Kyoichi Matsumoto, Junichi Takagi
  • Journal Title: Journal of Immunological Methods 352 Pages: 153-160
  • Peer Reviewed
• [Journal Article] Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation (2010)
  • Author(s): Norihisa Yasui, et al.
  • Journal Title: Journal of Immunological Methods 352 Pages: 153-160
  • Peer Reviewed
• [Journal Article] The role and structure of the carboxyl-terminal domain of the human voltage-gated proton channel Hv1. (2010)
  • Author(s): Shu Jie Li, et al.
  • Journal Title: The Journal of Biological Chemistry (In press)(掲載確定)
  • Peer Reviewed
• [Journal Article] Biological and immunological characteristics of hepatitis E virus-like particles based on the crystal structure. (2009)
  • Author(s): Tetsuo Yamashita, Yoshio Moria, Naoyuki Miyazaki, R. Holl, Cheng, Masato Yoshimura, Hideaki Unno, Ryoichi Shima, Kohji Moriishi, Tomitake Tsukihara, Tian Cheng Li, Naokazu Takeda, Tatsuo Miyamura, Yoshiharu Matsuura
  • Journal Title: Proc Natl Acad Sci U S A. 106(31) Pages: 12986-12991
  • Peer Reviewed
• [Journal Article] Effects of Ca2+ on Refolding of the Recombinant Hemolytic Lectin CEL-III in its Oligomerization and Hemolytic Abilities. (2009)
  • Author(s): Keigo Hisamatsu, Hideaki Unno, Shuichiro Goda, Tomomitsu Hatakeyama
  • Journal Title: Biosci. Biotechnol. Biochem. 73(5) Pages: 1203-1205
  • Peer Reviewed
• [Journal Article] New Role of Flavin as a General Acid-Base Catalyst with No Redox Function in Type 2 Isopentenyl Diphosphate Isomerase (2009)
  • Author(s): Hideaki Unno, Satoshi Yamashita, Yosuke Ikeda, Shin-ya Sekiguchi, Norie Yoshida, Tohru Yoshimura, Masami Kusunoki, Toru Nakayama, Tokuzo Nishino, Hisashi Hemmi
  • Journal Title: J. Biol. Chem. 284(14) Pages: 9160-9167
  • Peer Reviewed
• [Journal Article] Roles of the Valine Clusters in Domain 3 of the Hemolytic Lectin CEL-III in Its Oligomerization and Hemolytic Abilities (2009)
  • Author(s): Keigo Hisamatsu, Hideaki Unno, Shuichiro Goda, Tomomitsu Hatakeyama
  • Journal Title: Protein Pept. Lett. 16(4) Pages: 411-414
  • NAID: 120006984476
  • Peer Reviewed
• [Journal Article] Biological and immunological characteristics of hepatitis E virus-like particies based on the crystal structure. (2009)
  • Author(s): Tetsuo Yamashita, et al.
  • Journal Title: Proc Nafi Acad Sci U S A. 106(31) Pages: 12986-12991
  • Peer Reviewed
• [Journal Article] New Role of Flavin as a General Acid-Base Catalyst with No Redox Function in Type 2 Isopentenyl Diphosphate Isomerase (2009)
  • Author(s): Hideaki Unno, et al.
  • Journal Title: The Journal of Biological Chemistry 284(14) Pages: 9160-9167
  • Peer Reviewed
• [Journal Article] Roles of the Valine Clusters in Domain 3 of the Hemolytic Lectin CEL-III in Its Oligomerization and Hemolytic Abilities (2009)
  • Author(s): Keigo Hisamatsu, et al.
  • Journal Title: Protein Pept.Lett. 16(4) Pages: 411-414
  • Peer Reviewed
• [Journal Article] Flavin's new role as a general acid-base catalyst with no redox function in type 2 isonentenvl dinhosnhate isomerase (2009)
  • Author(s): Hideaki Unno
  • Journal Title: J. Biol. Chem. (掲載確定)(in press)
  • Peer Reviewed
• [Journal Article] Roles of the Valine Clusters in Domain 3 of the Hemolytic Lectin CEL-III in Its Olieomerization and Hemolytic Abilities (2009)
  • Author(s): Keigo Hisamatsu
  • Journal Title: Protein Pept. Lett. 16(in press)(掲載確定)
  • Peer Reviewed
• [Journal Article] Effects of Ca2+on Refolding of the Recombinant Hemolytic Lectin CEL-III (2009)
  • Author(s): Keigo Iiisamatsu
  • Journal Title: Biosci. Biotechnol. Biochem (掲載確定)(in press)
  • Peer Reviewed
• [Journal Article] Structural basis for substrate recognition and hydrolysis by mouse carnosinase CN2. (2008)
  • Author(s): Hideaki Unno, Tetsuo Yamashita, Sayuri Ujita, Nobuaki Okumura, Hiroto Otani, Akiko Okumura, Katsuya Nagai, Masami Kusunoki
  • Journal Title: J. Biol. Chem. 283(40) Pages: 27289-27299
  • Peer Reviewed
• [Journal Article] Crystal structure of the catalytic domain of Japanese encephalitis virus NS3 helicase/nucleoside triphosphatase at a resolution of 1. 8 A. (2008)
  • Author(s): Tetsuo Yamashita, Hideaki Unno, Yoshio Mori, Hideki Tani, Kohji Moriishi, Akihisa Takamizawa, Masanobu Agoh, Tomitake Tsukihara, Yoshiharu Matsuura
  • Journal Title: Virology 373(2) Pages: 426-436
  • Peer Reviewed
• [Journal Article] Structural basis for substrate recognition and hydrolysis by mouse carnosinase CN2 (2008)
  • Author(s): Hideaki Unno
  • Journal Title: J. Biol. Chem. 283(40) Pages: 27289-27299
  • Peer Reviewed
• [Journal Article] Crystal structure of the catalytic domain of Japanese encephalitis virus NS3 hplicase/nueleoside triphosohatase at a resolution of 1.8A (2008)
  • Author(s): Tetsuo Yamashita
  • Journal Title: Virology 373(2) Pages: 426-436
  • Peer Reviewed
• [Journal Article] Effects of Ca2+ on Refolding of the Recombinant Hemolytic Lectin CEL -III in its Oligomerization and Hemolytic Abilities
  • Author(s): Keigo Hisamatsu, et al.
  • Journal Title: Biosci.Biotechnol.Biochem. 73(5)2009 Pages: 1203-1205
  • Peer Reviewed
• [Presentation] タイプ2イソペンテニル二リン酸イソメラーゼの反応機構:一般酸塩基触媒としてのフラビンの機能 (2009)
  • Author(s): 邊見久
  • Organizer: 日本農芸化学会2009年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-29
• [Presentation] 樹状細胞表面受容体BDCA-2のC型レクチンドメインの大腸菌による発現 (2009)
  • Author(s): 佐知望美
  • Organizer: 日本農芸化学会2009年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] 光散乱法を用いたレクチン/糖含有ポリマー間相互作用の解析 (2009)
  • Author(s): 寺井康了
  • Organizer: 蛋白質と酵素の構造と機能に関する九州シンポジウム
  • Place of Presentation: 佐賀
• [Presentation] 溶血性レクチンCEL-IIIの自己会合ドメイン中のアミノ酸残基の役割 (2009)
  • Author(s): 久松啓伍
  • Organizer: 第9回日本蛋白質科学会年会
  • Place of Presentation: 熊本
• [Presentation] Structural and mutational studies of anthocyanin malonyltransferases establish the features of BAHD enzyme catalysis. (2008)
  • Author(s): Hideaki Unno
  • Organizer: Americal Crystallographic Association Meeting
  • Place of Presentation: アメリカ
  • Year and Date: 2008-06-03
• [Presentation] Structural and mutational studies of anthocyanin malonyltransferases stablish the features of BAHD enisTme catalysis (2008)
  • Author(s): Hideaki Unno
  • Organizer: Americal Crystallographie Association Meeting
  • Place of Presentation: アメリカテネシー州ノックスビル
  • Year and Date: 2008-06-03
• [Presentation] アントシアニンマロニルトランスフェラーゼのX線結晶構造解析 (2008)
  • Author(s): 海野英昭
  • Organizer: 日本生化学会九州支部例会
  • Place of Presentation: 福岡
  • Year and Date: 2008-05-17
• [Presentation] 光散乱法によるレクチン-糖鎖間相互作用解析 (2008)
  • Author(s): 畠山智充
  • Organizer: 平成20年度日本農芸化学会西日本支部大会
  • Place of Presentation: 長崎
• [Presentation] 溶血性レクチンCEL-III会合ドメインの部位特異的変異体作製とその性質 (2008)
  • Author(s): 久松啓伍
  • Organizer: 平成20年度日本農芸化学会西日本支部大会
  • Place of Presentation: 長崎
• [Presentation] グミ由来CEL-III膜貫通型複合体のX線結晶構造解析 (2008)
  • Author(s): 松本尚樹
  • Organizer: 平成20年度日本農芸化学会西日本支部大会
  • Place of Presentation: 長崎
• [Presentation] α-ガラクトシド特異的C型レクチンCEL-IVと糖との相互作用解析 (2008)
  • Author(s): 畠山智充
  • Organizer: 日本生化学会大会九州支部例会
  • Place of Presentation: 福岡