Molecules involved in intracellular trafficking of botulinum neurotoxin binding protein NTNHA

• Project/Area Number: 15K08474
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bacteriology (including mycology)
• Research Institution: Tokyo University of Agriculture
• Principal Investigator: Sagane Yoshimasa 東京農業大学, 生物産業学部, 教授 (00624660)
• Co-Investigator(Renkei-kenkyūsha): NIWA Koichi 東京農業大学, 生物産業学部, 教授 (20301012) ; MUTOH Shingo 鎌倉女子大学, 家政学部, 講師 (20749890) ; SUZUKI Tomonori 東京農業大学, 生命科学部, 准教授 (90453836)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2015: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: ボツリヌス食中毒 / 細胞内輸送 / 経口毒素 / ボツリヌス毒素 / アクチンフィラメント / 抗体 / 非毒非血球凝集素 / ミオシン

Outline of Final Research Achievements

Nontoxic nonhemagglutinin (NTNHA) is a protein that binds to botulinum neurotoxin (BoNT), a causative agent of botulism food poisoning. NTNHA is known to increase oral toxicity of toxin by protecting BoNT from digestive juices, but its function is remained unclear. This study aims to elucidate the molecules involved in intracellular transport of NTNHA. The results of this study suggested that "actin-myosin" system is involved in NTNHA intracellular transport. Also, vesicles involved in the intracellular transport of NTNHA were revealed. The results of this study are expected to greatly contribute to the elucidation of the invasion mechanism of botulinum toxin into the body.

Research Products

• [Journal Article] Building-block architecture of botulinum toxin complex: Conformational changes provide insights into the hemagglutination ability of the complex (2017)
  • Author(s): Suzuki T, Sagane Y, Matsumoto T, Hasegawa K, Yamano A, Niwa K, Watanabe T
  • Journal Title: Biochemistry and Biophysics Reports Volume: 9 Pages: 67-71
  • DOI: 10.1016/j.bbrep.2016.11.008
  • Peer Reviewed
• [Journal Article] Reversible Association of the Hemagglutinin Subcomplex, HA-33/HA-17 Trimer, with the Botulinum Toxin Complex (2017)
  • Author(s): Sagane Y, Mutoh S, Koizumi R, Suzuki T, Miyashita S, Miyata K, Ohyama T, Niwa K, Watanabe T
  • Journal Title: The Protein Journal Volume: 36 Issue: 5 Pages: 417-424
  • DOI: 10.1007/s10930-017-9733-y
  • Peer Reviewed
• [Journal Article] Construction of ‘‘Toxin Complex’’ in a Mutant Serotype C Strain of Clostridium botulinum Harboring a Defective Neurotoxin Gene (2016)
  • Author(s): Tomonori Suzuki, Thomas Nagano, Koichi Niwa, Masataka Uchino, Motohiro Tomizawa, Yoshimasa Sagane, and Toshihiro Watanabe
  • Journal Title: Current Microbiology Volume: 74 Issue: 1 Pages: 49-54
  • DOI: 10.1007/s00284-016-1150-5
  • Peer Reviewed
• [Journal Article] "Non-Toxic" Proteins of the Botulinum Toxin Complex Exert In-vivo Toxicity. (2016)
  • Author(s): Shin-Ichiro Miyashita, Yoshimasa Sagane, Tomonori Suzuki, Takashi Matsumoto, Koichi Niwa, and Toshihiro Watanabe
  • Journal Title: Scientific Reports Volume: 6 Issue: 1
  • DOI: 10.1038/srep31043
  • Peer Reviewed / Open Access
• [Journal Article] Conformational divergence in the HA-33/HA-17 trimer of serotype C and D botulinum toxin complex. (2016)
  • Author(s): Yoshimasa Sagane, Shintaro Hayashi, Tomonori Akiyama, Takashi Matsumoto, Kimiko Hasegawa, Akihito Yamano, Tomonori Suzuki, Koichi Niwa, Toshihiro Watanabe, and Shunsuke Yajima
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 476 Issue: 4 Pages: 280-285
  • DOI: 10.1016/j.bbrc.2016.05.113
  • Peer Reviewed
• [Journal Article] Data describing inhibitory profiles of sugars against hemagglutination by the botulinum toxin complex of Clostridium botulinum serotypes C and D (2016)
  • Author(s): Yoshimasa Sagane, Minoru Ito, Keita Miyata, Tomonori Suzuki, Koichi Niwa, Suguru Oguri, and Toshihiro Watanabe
  • Journal Title: Data in Brief Volume: 9 Pages: 413-416
  • DOI: 10.1016/j.dib.2016.09.014
  • Peer Reviewed / Open Access
• [Journal Article] Isolation of botulinolysin, a thiol-activated hemolysin, from serotype D Clostridium botulinum: A species-specific gene duplication in Clostridia (2016)
  • Author(s): Tomonori Suzuki, Thomas Nagano, Koichi Niwa, Shingo Mutoh, Masataka Uchino, Motohiro Tomizawa, Yoshimasa Sagane, and Toshihiro Watanabe
  • Journal Title: Microbiological Research Volume: 193 Pages: 20-29
  • DOI: 10.1016/j.micres.2016.08.011
  • Peer Reviewed
• [Journal Article] Hemagglutinin Gene Shuffling among Clostridium botulinumSerotypes C and D Yields Distinct Sugar Recognition of the Botulinum Toxin Complex (2015)
  • Author(s): Miyata K, Suzuki T, Hayashi S, Miyashita S, Ohyama T, Niwa K, Watanabe T, Sagane Y
  • Journal Title: FEMS Pathogens and Disease Volume: 73 Issue: 7 Pages: ftv054-ftv054
  • DOI: 10.1093/femspd/ftv054
  • Peer Reviewed
• [Presentation] Internalization and intercellular trafficking of the serotype D botulinum neurotoxin binding protein, nontoxic nonhemagglutinin, in epithelial and endothelial cells. (2017)
  • Author(s): Sagane, Y., Miyashita, S. -I., Hosoya, K., Karatsu, S., Kurihara, A., Sugawara, C., Niwa, K., Watanabe, T.
  • Organizer: 10th International Conference on the Molecular Biology and Pathogenesis of the Clostridia.
  • Int'l Joint Research
• [Presentation] “Non-toxic” proteins of the botulinum toxin complex exert cytotoxicity with vacuole formation in the cell. (2017)
  • Author(s): Sagane, Y., Miyashita, S. -I., Kurihara, A., Sugawara, C., Hosoya, K., Karatsu, S., Niwa, K., Watanabe, T.
  • Organizer: 10th International Conference on the Molecular Biology and Pathogenesis of the Clostridia.
  • Int'l Joint Research