Combined analysis of tertiary structure and function(s) of Clostridium botulinum type C and D phospholipase C

2010 Fiscal Year Final Research Report

• Project/Area Number: 21790421
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Bacteriology (including Mycology)
• Research Institution: University of Miyazaki (2010); Okayama University (2009)
• Principal Investigator: SAKAGUCHI Yoshihiko University of Miyazaki, IR推進機構, 助教 (70403491)
• Project Period (FY): 2009 – 2010
• Keywords: ボツリヌス菌 / ホスホリパーゼC / ウエルシュ菌α毒素 / 卵黄活性 / 溶血活性 / ホスファチジルコリン / スフィンゴミエリン / リポソーム

Research Abstract

Clostridium botulinum cultures are classified into seven types, types A to G, based on the antigenicity of the neurotoxins produced. Of these seven types, only types C and D produce phospholipase C (lecithinase, PLC) in addition to the neurotoxin. Phospholipase of C. perfringens is known as α-toxin, which is the best characterized of all clostridial phospholipases and is an important agent of gas gangrene. The toxin, which exhibits phospholipase C and sphingomyelinase (SMase) activities, causes hemolysis, necrosis, and death. Here we determined the PLC genes of C. botulinum type C and D (Cbotα and Dbotα) by using inverse PCR and primer walking, and expressed as a GST-fusion protein. We compare the enzymatic and biological activities (egg-yolk activity, hemolysis, and mouse lethality) of the purified protein of C. perfringens α-toxin, Cbotα, and Dbotα.

Research Products

• [Journal Article] A new cytosolicpathway from a Parkinsondisease-associated kinase, BRPK/PINK1:activation of AKT via MTORC2. (2011)
  • Author(s): Murata, H., Sakaguchi, M., Jin, Y., Sakaguchi, Y., Futami, J., Yamada, H., Kataoka, K., Huh, NH.
  • Journal Title: J.Biol.Chem. 286(9) Pages: 7182-7189
  • Peer Reviewed
• [Journal Article] Transcriptionalregulation of a brown adipocyte-specificgene, UCP1, by KLF11 and KLF15. (2010)
  • Author(s): Yamamoto, K., Sakaguchi, M., Medina, RJ., Niida A., Sakaguchi, Y., Miyazaki, M., Kataoka, K., Huh, NH.
  • Journal Title: Biochem.Biophys.Res.Commun. 400(1) Pages: 175-180
  • Peer Reviewed
• [Presentation] ボツリヌス菌ホスホリパーゼCの酵素活性と生物活性, BMB2010 (2010)
  • Author(s): 阪口義彦, 小田真隆, Ni Nengah DwiFatmawati, 山本由弥子, 唐澤忠宏, 小林敬子, 永浜政博, 櫻井純, 小熊惠二
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド(神戸)
  • Year and Date: 20101207-20101210
• [Presentation] 多機能受容体RAGEの下流信号伝達機構の解明, BMB2010 (2010)
  • Author(s): 阪口政清, 村田等, 山本健一, 小野智之, 阪口義彦, 本山晃, 日比野利彦, 片岡健, 許南浩
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド(神戸)
  • Year and Date: 20101207-20101210
• [Presentation] A型ボツリヌス神経毒素の三叉神経節培養細胞への結合と取り込み (2010)
  • Author(s): 山本由弥子, 張凱, Ni Nengah DwiFatmawati, 申蓮花, 馬少博, 阪口義彦, 鈴木智典, 小熊惠二
  • Organizer: 第63回日本細菌学会中国・四国支部総会
  • Place of Presentation: 松山大学(松山市)
  • Year and Date: 20101016-20101017
• [Presentation] C型とD型ボツリヌス毒素を支配するバクテリオファージのゲノム解析 (2010)
  • Author(s): 阪口義彦
  • Organizer: 第3回ファージ研究会
  • Place of Presentation: 大阪大学蛋白質研究所(大阪)
  • Year and Date: 20100909-20100910
• [Presentation] ボツリヌス菌ホスホリパーゼCの酵素活性と生物活性 (2010)
  • Author(s): 阪口義彦, 小田真隆, Ni Nengah DwiFatmawati, 山本由弥子, 難波ひかる, 唐澤忠宏, 小林敬子, 永浜政博, 櫻井純, 小熊惠二
  • Organizer: 第57回トキシンシンポジウム
  • Place of Presentation: 長浜ロイヤルホテル(滋賀)<招待講演>
  • Year and Date: 20100714-20100716
• [Remarks] ホームページ等