Elucidation of the structure and function of a photoactivated adenylyl cyclase from a sulfer bacterium

• Project/Area Number: 25440072
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: Toho University
• Principal Investigator: ISEKI Mineo 東邦大学, 薬学部, 准教授 (60414009)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 光センサー / フラビン / アデニル酸シクラーゼ / cAMP / 光生物学 / 光受容体

Outline of Final Research Achievements

This research is aimed at elucidating the activation mechanism of photoactivated adenylyl cyclase (PAC). We found a new homolog of PAC in the genome sequence of a cyanobacterium Oscillatoria acuminata and demonstrated that the homolog (OaPAC) can be easily expressed and purified from E. coli as soluble form. Using the purified protein, we successfully determined the photochemical and biochemical properties of the protein, and solved its crystal structure under collaboration with structure biologists.

Research Products

• [Journal Article] Photoactivated adenylyl cyclase (PAC) reveals novel mechanisms underlying cAMP-dependent axonal morphogenesis (2016)
  • Author(s): Zhou Z, Tanaka KF, Matsunaga S, Iseki M, Watanabe M, Matsuki N, Ikegaya Y, Koyama R
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 19679-19679
  • DOI: 10.1038/srep19679
  • Peer Reviewed / Open Access
• [Journal Article] A simple method to prepare a biopterin sensor chip applicable to crude samples (2015)
  • Author(s): Junko Takeda, Kanako Harada, Chisae Ishitoko, Hiroshi Ueno, Mineo Iseki, Masakatsu Watanabe
  • Journal Title: J. Biol. Macromol. Volume: 15 Pages: 29-39
  • NAID: 130007494422
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Possible involvement of a tetrahydrobiopterin in photoreception for UV-B-induced anthocyanin synthesis in carrot. (2014)
  • Author(s): Takeda J., Nakata R., Ueno H., Murakami A., Iseki M. and Watanabe M.
  • Journal Title: Photochemistry and Photobiology Volume: 90 Issue: 5 Pages: 1043-1049
  • DOI: 10.1111/php.12302
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Blue light regulates the rhythm of diurnal vertical migration in the raphidophyte red-tide alga Chattonella antiqua. (2013)
  • Author(s): Tomoyuki Shikata, Shigeru Matsunaga, Mineo Iseki, Hiroyo Nishide, Sho-Ichi Higashi,Yasuhiro Kamei, Mineo Yamaguchi, Ian R. Jenkinson, Masakatsu Watanabe
  • Journal Title: J. Plankton Res. Volume: 35 Issue: 3 Pages: 542-552
  • DOI: 10.1093/plankt/fbt006
  • Peer Reviewed
• [Journal Article] Identification of photoactivated adenylyl cyclases in <i>Naegleria australiensis</i> and BLUF-containing protein in <i>Naegleria fowleri</i> (2013)
  • Author(s): Yasukawa H, Sato A, Kita A, Kodaira K, Iseki M, Takahashi T, Shibusawa M, Watanabe M, Yagita K.
  • Journal Title: The Journal of General and Applied Microbiology Volume: 59 Issue: 5 Pages: 361-369
  • DOI: 10.2323/jgam.59.361
  • NAID: 130004449379
  • ISSN: 0022-1260, 1349-8037
  • Peer Reviewed
• [Presentation] Euglena gracilisのステップダウン光驚動反応に関与する光センサータンパク質の探索 (2015)
  • Author(s): 田上真也, 石川孝博, 伊関峰生
  • Organizer: ユーグレナ研究会第31回研究集会
  • Place of Presentation: Kitenビル コンベンションホール（宮崎県宮崎市）
  • Year and Date: 2015-11-07
• [Presentation] 光活性化アデニル酸シクラーゼ合成酵素PACの活性化機構解明 (2015)
  • Author(s): 大木 規央, 杉山 佳奈子, 河合 文啓, 松永 茂, 柴山 修哉, 伊関 峰生, 朴 三用
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学角間キャンパス（石川県金沢市）
  • Year and Date: 2015-09-15
• [Presentation] MALDI-TOF-MSを用いたBLUFドメインの同位体標識の解析 (2015)
  • Author(s): 永井 貴士, 岩田 達也, 伊藤 奨太, 伊関 峰生, 渡辺 正勝, 北川 慎也, 神取 秀樹
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学角間キャンパス（石川県金沢市）
  • Year and Date: 2015-09-14
• [Presentation] シアノバクテリア由来の新規光活性化アデニル酸シクラーゼ (2015)
  • Author(s): 伊関 峰生, 二瓶 悠樹, 田中 瞳, 大木 規央, 杉山 佳奈子, 河合 文啓, 松永 茂, 高橋 哲郎, 朴 三用
  • Organizer: 日本植物学会第79回大会
  • Place of Presentation: 朱鷺メッセ：新潟コンベンションセンター（新潟県新潟市）
  • Year and Date: 2015-09-06
• [Presentation] 光活性化アデニル酸シクラーゼの構造と機能－生体機能光操作への応用を目指して－ (2015)
  • Author(s): 伊関峰生
  • Organizer: 第57 回日本放線菌学会学術講演会
  • Place of Presentation: 微生物化学研究所（東京都品川区）
  • Year and Date: 2015-02-27
  • Invited
• [Presentation] ミドリムシにおける光運動制御マシナリーの解明 (2014)
  • Author(s): 岩崎憲治、宮崎直幸、伊関峰生、長谷川浩司、成田哲博、松永茂、建部益美、上村慎治、、渡辺正勝
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2014-09-25
• [Presentation] cAMPによる軸索分枝と伸長の独立制御 (2014)
  • Author(s): 周 至文，田中 謙二，松永 茂，伊関 峰生，渡辺 正勝，松木 則夫，池谷 裕二，小山 隆太
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-09-13
• [Presentation] Analysis of intramolecular signal transduction in the photoactivated adenylyl cyclase of the sulfur bacterium Beggiatoa sp (2014)
  • Author(s): Mineo Iseki, Shoko Uekusa, Asami Yamagata, Koji Hasegawa, Hiro Yasukawa, Tetsuo Takahashi, Masakatsu Watanabe
  • Organizer: 16th International Congress of Photobiology
  • Place of Presentation: Universidad Nacional de Cordoba, Cordoba, Argentina
  • Year and Date: 2014-09-09
• [Presentation] in vivo (in cell) crystal structural analysis using EM
  • Author(s): Naoyuki Miyazaki, Iseki Mineo, Koji Hasegawa, Akihiro Narita, Shinichi Adachi, Watanabe masakatsu, Kenji Iwasaki
  • Organizer: Structural Life Science 7th International Conference on Structural Genomics
  • Place of Presentation: 京王プラザホテル札幌（北海道札幌市）
• [Presentation] ミドリムシ鞭毛由来青色蛍光タンパク質の機能解析
  • Author(s): 齊藤佑樹, 堀池裕一郎, 川上 亮, 松永 茂, 渡辺正勝, 高橋哲郎, 伊関峰生
  • Organizer: 第29回ユーグレナ研究会
  • Place of Presentation: 筑波大学（茨城県つくば市）
• [Book] Optogenetics: Light-Sensing Proteins and Their Applications (2015)
  • Author(s): Mineo Iseki, Tetsuo Takahashi
  • Total Pages: 409
  • Publisher: Springer Japan
• [Book] 光化学の事典 (2014)
  • Author(s): 伊関峰生
  • Total Pages: 436
  • Publisher: 朝倉書店