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Cell-layer transport mechanism of botulinum neurotoxin-binding protein, nontoxic nonhemagglutinin

Research Project

Project/Area Number 25860325
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Bacteriology (including mycology)
Research InstitutionTokyo University of Agriculture

Principal Investigator

SAGANE Yoshimasa  東京農業大学, 生物産業学部, 教授 (00624660)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywordsボツリヌス食中毒 / 体内輸送 / 経口毒素 / タンパク質毒素 / 消化管輸送
Outline of Final Research Achievements

Botulinum toxin (BoNT) complex (TC) traverses the gastrointestinal tract and circulation, eventually becoming localized in neuromuscular junctions. During this process, BoNT must pass through cells, thus from the intestinal lumen to the cells of the intestinal tract and blood vessels. The botulinum TC is formed by association of the BoNT with at least one nontoxic protein, nontoxic nonhemagglutinin (NTNHA). In the present work, we examined the binding and transcytosis of serotype D NTNHA protein in epithelial and endothelial cells to clarify the role played by the protein in toxin delivery. Our studies showed that NTNHA bound to and transcytosed across IEC-6 and BAEC cells. While NTNHA also bound to MDCK or Caco-2 cells, but did not traverse across MDCK or Caco-2 cells. Such specificity of NTNHA protein transcytosis may explain why only some animals are sensitive to botulinum toxin. The sensitivity depends on the toxin serotype in play, and the route of toxin delivery.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • Research Products

    (14 results)

All 2015 2014 2013

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Open Access: 1 results,  Acknowledgement Compliant: 2 results) Presentation (7 results)

  • [Journal Article] 87. Purification, crystallization and preliminary X-ray analysis of a HA17-HA70 (HA2-HA3) complex from Clostridium botulinum type C progenitor toxin.2014

    • Author(s)
      Chikako Iwasa, Takashi Tonozuka, Masaya Shinoda, Yoshimasa Sagane, Koichi Niwa, Toshihiro Watanabe, Hiromi Yoshida, Shigehiro Kamitori, Toshifumi Takao, Keiji Oguma, Atsushi Nishikawa
    • Journal Title

      Acta Crystallographica

      Volume: F70 Issue: 1 Pages: 64-67

    • DOI

      10.1107/s2053230x13032378

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization and preliminary X-ray analysis of a novel haemagglutinin component of the toxin complex of serotype C Clostridium botulinum.2014

    • Author(s)
      Shintaro Hayashi, Tomonori Akiyama, Yoshimasa Sagane, Shin-Ichiro Miyashita, Toshihiro Watanabe, Shunsuke Yajima, and Koichi Niwa
    • Journal Title

      Acta Crystallography F Structural Biology Communication

      Volume: 70 Issue: 3 Pages: 370-373

    • DOI

      10.1107/s2053230x14003094

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of the interaction region between hemagglutinin components of the botulinum toxin complex2014

    • Author(s)
      Suzuki T, Miyashita S, Hayashi S, Miyata K, Inui K, Kondo Y, Miyazaki S, Ohyama T, Niwa K, Watanabe T, Sagane Y
    • Journal Title

      International Journal of Biological Macromolecules

      Volume: 65 Pages: 284-288

    • DOI

      10.1016/j.ijbiomac.2014.01.052

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Host-cell specificity and transcytosis of nontoxic nonhemagglutinin protein of botulinum neurotoxin serotype D2014

    • Author(s)
      Shin-Ichiro Miyashita, Koichi Niwa, Toshihiro Watanabe, and Yoshimasa Sagane
    • Journal Title

      FEMS Microbiology Letter

      Volume: 357 Pages: 115-122

    • DOI

      10.1111/1574-6968.12527

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Transport of the botulinum neurotoxin-associating protein, nontoxic nonhemagglutinin, across the rat small intestinal epithelial cell monolayer2013

    • Author(s)
      Shin-Ichiro Miyashita, Yoshimasa Sagane, Koichi Niwa, Toshihiro Watanabe
    • Journal Title

      FEMS Microbiology Letter

      Volume: 346 Issue: 1 Pages: 73-80

    • DOI

      10.1111/1574-6968.12205

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Sugar-induced conformational change found in the HA-33/HA-17 trimer of the botulinum toxin complex2013

    • Author(s)
      Yoshimasa Sagane, Shintaro Hayash, Takashi Matsumoto, Shin-ichro Miyashita, Ken Inui, Keita Miyata, Shunsuke Yajima, Tomonori Suzuki, Kimiko Hasegawa, Akihito Yamano, Atushi Nishikawa, Tohru Ohyama, Toshihiro Watanabe, Koichi Niwa
    • Journal Title

      Biochem. Biophys. Res. Commun

      Volume: 438 Issue: 3 Pages: 483-487

    • DOI

      10.1016/j.bbrc.2013.07.112

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Botulinum Toxin Complex Increases Paracellular Permeability in Intestinal Epithelial Cells via Activation of p38 Mitogen-Activated Protein Kinase2013

    • Author(s)
      Miyashita S-I., Sagane Y., Inui K., Hayashi S., Miyata K., Suzuki T., Ohyama T., Watanabe T., Niwa K.
    • Journal Title

      Journal of Veterinary Medical Science

      Volume: 75 Issue: 12 Pages: 1637-1642

    • DOI

      10.1292/jvms.13-0164

    • NAID

      130003362264

    • ISSN
      0916-7250, 1347-7439
    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] ボツリヌスC 型菌Yoichi 株の産生するHA-33/HA-17 複合体の構造解析2015

    • Author(s)
      林 慎太郎,相根 義昌,松本 崇,秋山 友了,宮下 慎一郎, 武藤 信吾,山野 昭人,矢嶋 俊介,渡部 俊弘,丹羽 光一
    • Organizer
      第88回日本細菌学会総会
    • Place of Presentation
      岐阜
    • Year and Date
      2015-03-26 – 2015-03-28
    • Related Report
      2014 Annual Research Report
  • [Presentation] ボツリヌスC型菌Yoichi株が産生するHA-33の構造解析2014

    • Author(s)
      林 慎太郎, 相根 義昌, 松本 崇, 秋山 友了, 宮下 慎一郎, 長谷川 仁子, 山野 昭人, 矢嶋 俊介, 渡部 俊弘, 丹羽 光一
    • Organizer
      第87回日本生化学会大会
    • Place of Presentation
      京都
    • Year and Date
      2014-10-15 – 2014-10-18
    • Related Report
      2014 Annual Research Report
  • [Presentation] D型ボツリヌス神経毒素結合タンパク質 非毒非血球凝集素の細胞種特異的なトランスサイトーシス2014

    • Author(s)
      宮下 慎一郎, 林 慎太郎, 丹羽 光一, 渡部 俊弘, 相根 義昌
    • Organizer
      第87回日本生化学会大会
    • Place of Presentation
      京都
    • Year and Date
      2014-10-15 – 2014-10-18
    • Related Report
      2014 Annual Research Report
  • [Presentation] D型ボツリヌス神経毒素結合タンパク質非毒非血球凝集素のヒト大腸癌細胞への結合および透過2014

    • Author(s)
      宮下慎一郎,相根義昌,林慎太郎,鈴木智典,丹羽光一,渡部俊弘
    • Organizer
      第87回日本細菌学会総会
    • Place of Presentation
      東京
    • Related Report
      2013 Research-status Report
  • [Presentation] ボツリヌスC型菌Yoichi株の産生するHA-33/HA-17複合体の構造2014

    • Author(s)
      林慎太郎, 相根義昌,松本崇, 秋山友了,宮下慎一郎,長谷川仁子,山野昭人,矢嶋俊介, 渡部俊弘,丹羽光一
    • Organizer
      第87回日本細菌学会総会
    • Place of Presentation
      東京
    • Related Report
      2013 Research-status Report
  • [Presentation] D型ボツリヌス神経毒素結合タンパク質 NTNHAはラット小腸上皮細胞へ結合し細胞層を透過する.2013

    • Author(s)
      宮下慎一郎, 相根義昌, 林慎太郎, 鈴木智典,丹羽光一, 渡部俊弘.
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report
  • [Presentation] ボツリヌスC型菌Yoichi株が産生するHA-33の結晶化2013

    • Author(s)
      林慎太郎, 相根義昌, 秋山友了, 宮下慎一郎, 鈴木智典,矢嶋俊介, 渡部俊弘, 丹羽光一.
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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