Investigation of PE as a novel receptor for botulinum neurotoxin type D

• Project/Area Number: 21790430
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Bacteriology (including Mycology)
• Research Institution: Fujita Health University
• Principal Investigator: TSUKAMOTO Kentaro 藤田保健衛生大学, 医学部, 講師 (80434596)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 病原性 / 細菌毒素 / 細菌 / 蛋白質 / P19細胞 / ボツリヌス神経毒素 / 受容体 / ガングリオシド / ボツリヌス菌 / 毒素受容体

Research Abstract

Botulinum neurotoxin type D possesses binding activity to phosphatidylethanolamine(PE), however, it is not unclear whether PE could mediate the internalization of the toxin into neuronal cells. In this study, we determined crystal structure of the receptor binding domain of the toxin. Moreover, we found that mouse embryonic carcinoma P19 cells exhibited high sensitivity to botulinum neurotoxin type C and D/C mosaic toxin, thus making them a novel susceptible cell line for research into botulinum neurotoxins.

Research Products

• [Journal Article] Structural and mutational analyses of the receptor binding domain of botulinum D/C mosaic neurotoxin : Insight into the ganglioside binding mechanism (2011)
  • Author(s): Nuemket N., Tanaka Y., Tsukamoto K., Tsuji T., Nakamura K., Kozaki S., Yao M., Tanaka I.
  • Journal Title: Biochem Biophys Res Commun Volume: Vol.411 Pages: 433-9
  • Peer Reviewed
• [Journal Article] Single chain variable fragment antibodies against Shiga toxins isolated from a human antibody phage display library (2011)
  • Author(s): Neri P., Shigemori N., Hamada-Tsutsumi S., Tsukamoto K., Arimitsu H., Shimizu T., Akahori Y., Kurosawa Y., Tsuji T.
  • Journal Title: Vaccine Volume: Vol.29 Pages: 5340-6
  • Peer Reviewed
• [Journal Article] Structural and mutational analyses of the receptor binding domain of botulinum D/C mosaic neurotoxin : Insight into the ganglioside binding mechanism (2011)
  • Author(s): Nipawan Nuemket, et.al.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 411 Pages: 433-439
  • Peer Reviewed
• [Journal Article] Single chain variable fragment antibodies against Shiga toxins isolated from a human antibody phage display library (2011)
  • Author(s): Paola Neri, et al
  • Journal Title: Vaccine Volume: 29 Pages: 5340-5346
  • Peer Reviewed
• [Journal Article] Preliminary X-ray crystallographic study of the receptor-binding domain of the D/C mosaic neurotoxin from Clostridium botulinum (2010)
  • Author(s): Nuemket N., Tanaka Y., Tsukamoto K., Tsuji T., Nakamura K., Kozaki S., Yao M., Tanaka I.
  • Journal Title: Acta Crystallogr Sect F Struct Biol Cryst Commun Volume: Vol.66 Pages: 608-10
  • Peer Reviewed
• [Journal Article] Preliminary X-ray crystallographic study of the receptor-binding domain of the D/C mosaic neurotoxin from Clostridium botulinum (2010)
  • Author(s): Nuemket, N., et.al
  • Journal Title: Acta Crystallogr Sect F Struct Biol Cryst Commun Volume: 66 Pages: 608-610
  • Peer Reviewed
• [Journal Article] Nucleotide sequence analysis of the enterotoxigenic Escherichia coli Ent plasmid (2009)
  • Author(s): Ochi S., Shimizu T., Ohtani K., Ichinose Y., Arimitsu H., Tsukamoto K., Kato M., Tsuji T.
  • Journal Title: DNA Res Volume: Vol.16 Pages: 299-309
  • Peer Reviewed
• [Journal Article] Lincomycin-induced over-expression of mature recombinant cholera toxin B subunit and the holotoxin in Escherichia coli (2009)
  • Author(s): Arimitsu H., Tsukamoto K., Ochi S., Sasaki K., Kato M., Taniguchi K., Oguma K., Tsuji T.
  • Journal Title: Protein Expr Purif Volume: Vol.67 Pages: 96-103
  • Peer Reviewed
• [Journal Article] Lincomycin-induced over-expression of mature recombinant cholera toxin B subunit and the holotoxin in Escherichia coli (2009)
  • Author(s): Arimitsu, H.et al.
  • Journal Title: Protein Expression Purification 67 Pages: 96-103
  • Peer Reviewed
• [Journal Article] Nucleotide sequence analysis of the enterotoxigenic Escherichia coli Entplasmid (2009)
  • Author(s): Ochi, S.et al.
  • Journal Title: DNA Research 16 Pages: 299-309
  • Peer Reviewed
• [Presentation] 胚性腫瘍細胞P19を用いたボツリヌス神経毒素の作用機構の解析 (2012)
  • Author(s): 塚本健太郎
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎県長崎市
  • Year and Date: 2012-03-29
• [Presentation] 胚性腫瘍細胞P19を用いたボツリヌス神経毒素の作用機構の解析 (2012)
  • Author(s): 塚本健太郎
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎ブリックホール(長崎県)
  • Year and Date: 2012-03-29
• [Presentation] Structure and activity of receptor binding domain of Clostridium botulinum D/C mosaic neurotoxin (2011)
  • Author(s): Kentaro Tsukamoto
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: Sapporo, Japan
  • Year and Date: 2011-09-07
• [Presentation] Structure and activity of receptor binding domain of Clostridium botulinum D/C mosaic neurotoxin (2011)
  • Author(s): Kentaro Tsukamoto
  • Organizer: International Union of Microbiological Societies 2011 Congress
  • Place of Presentation: 札幌コンベンションセンター(北海道)(招待講演)
  • Year and Date: 2011-09-07
• [Presentation] ボツリヌスC型及びDCモザイク神経毒素は異なるガングリオシドを介して細胞内に侵入する (2010)
  • Author(s): 西脇啓太
  • Organizer: 第47回日本細菌学会中部支部総会
  • Place of Presentation: 新潟県新潟市
  • Year and Date: 2010-10-23
• [Presentation] ボツリヌスC型およびDCモザイク神経毒素は異なるガングリオシドを介して細胞内に侵入する (2010)
  • Author(s): 西脇啓太, 他
  • Organizer: 第47回日本細菌学会中部支部総会
  • Place of Presentation: 新潟県新潟市
  • Year and Date: 2010-10-23
• [Presentation] ボツリヌス神経毒素受容体結合領域の結晶構造と細胞内侵入機構の解析 (2010)
  • Author(s): 塚本健太郎
  • Organizer: 第57回トキシンシンポジウム
  • Place of Presentation: 滋賀県長浜市
  • Year and Date: 2010-07-15
• [Presentation] ボツリヌスDCモザイク毒素重鎖C末端領域の構造と活性について (2010)
  • Author(s): 塚本健太郎
  • Organizer: 第8回感染症沖縄フォーラム
  • Place of Presentation: 沖縄県宜野湾市
  • Year and Date: 2010-02-12
• [Presentation] EC細胞を用いたボツリヌス神経毒素重鎖C末端領域の機能解析 (2009)
  • Author(s): 塚本健太郎
  • Organizer: 第46回日本細菌学会中部支部総会
  • Place of Presentation: 愛知県名古屋市
  • Year and Date: 2009-10-23
• [Presentation] ボツリヌス神経毒素の脂質受容体に対する結合活性の解析 (2009)
  • Author(s): 塚本健太郎
  • Organizer: 第41回藤田学園医学会
  • Place of Presentation: 愛知県豊明市
  • Year and Date: 2009-10-02
• [Presentation] ボツリヌス神経毒素重鎖C末端領域の脂質受容体に対する結合活性 (2009)
  • Author(s): 塚本健太郎
  • Organizer: 第56回トキシンシンポジウム
  • Place of Presentation: 岐阜県岐阜市
  • Year and Date: 2009-08-26