Purification and identification of a novel PTTH receptor in Bombyx mori

• Project/Area Number: 26660266
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Insect science
• Research Institution: Aichi Gakuin University (2015-2016); Nagoya University (2014)
• Principal Investigator: Mizoguchi Akira 愛知学院大学, 教養部, 教授 (60183109)
• Co-Investigator(Kenkyū-buntansha): 片岡 宏誌 東京大学, 大学院新領域創成科学研究科, 教授 (60202008)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 昆虫 / 前胸腺刺激ホルモン / 受容体 / カイコガ / 精製

Outline of Final Research Achievements

A putative PTTH receptor on the plasma membrane of the silkmoth prothoracic glands (PGs) was detected by Western blotting. The conjugate of this protein with PTTH had Mr of 120kDa and was not detected in any tissues other than the PGs. To purify this protein, the PGs were incubated with His-tagged PTTH, and the PTTH and receptor were linked to each other with a cross-linker, followed by solubilization with detergent. The resultant conjugate was successfully affinity-purified using magnetic beads for His-tag isolation, although the purity was still insufficient for sequence analysis. We tried to use immunoprecipitation for further purifying the PTTH-receptor conjugate without success.

Research Products

• [Journal Article] Myosuppression is involved in the regulation of pupal diapause in the cabbage army moth Mamestra brassicae.7 (2017)
  • Author(s): Yamada, N., Kataoka, H., Mizoguchi, A.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 41651-41651
  • DOI: 10.1038/srep41651
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Rab proteins in the brain and corpus allatum of Bombyx mori (2016)
  • Author(s): Uno.T., Furutani,M.,Watanabe,C., Sakamoto.K., Uno,Y., Kanamaru,K.,
  • Journal Title: Histochemistry and Cell Biology Volume: 未定 Issue: 1 Pages: 59-69
  • DOI: 10.1007/s00418-016-1422-y
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Endocrine Mechanisms Regulating Post-Diapause Development in the Cabbage Armyworm, Mamestra brassicae. (2016)
  • Author(s): Yamada, N., Okamoto, N., Kataoka, H., Mizoguchi, A.
  • Journal Title: PLos One Volume: 11 Issue: 1 Pages: e0146619-e0146619
  • DOI: 10.1371/journal.pone.0146619
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Isoform-specific expression of the neuropeptide orcokinin in Drosophila melanogaster. (2015)
  • Author(s): Chen, J., Choi, M.S., Mizoguchi, A., Veenstra, J.A., Kang, K., Kim, Y.-J., Kwon, J.Y.
  • Journal Title: Peptides Volume: 68 Pages: 50-57
  • DOI: 10.1016/j.peptides.2015.01.002
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Disruption of diapause induction by TALEN-based gene mutagenesis in relation to a unique neuropeptide signaling pathway in Bombyx. (2015)
  • Author(s): Shiomi, K., Takasu, Y., Kunii, M., Tsuchiya, R., Mukaida, M., Kobayashi, M., Sezutsu, H., Ichida, M., Mizoguchi, A.
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 15566-15566
  • DOI: 10.1038/srep15566
  • NAID: 120007101413
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 蛹休眠の調節機構 ―ヨトウガの蛹休眠を中心に－. (2015)
  • Author(s): 溝口明
  • Journal Title: 蚕糸・昆虫バイオテック Volume: 84 Pages: 135-143
  • NAID: 130006162018
• [Journal Article] Positive feedback regulation of prothoracicotropic hormone secretion by ecdysteroid - A mechanism that determines the timing of metamorphosis. (2015)
  • Author(s): 57.Mizoguchi, A., Kamimura, M., Kiuchi, M., Kataoka, H.
  • Journal Title: Insect Biochem. Mol. Biol. 58, 39-45. Volume: 58 Pages: 39-45
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Isoform-specific expression of the neuropeptide orcokinin in Drosophila melanogaster. (2015)
  • Author(s): 58.Chen, J., Choi, M.S., Mizoguchi, A., Veenstra, J.A., Kang, K., Kim, Y.-J., Kwon, J.Y.
  • Journal Title: Peptides Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Orcokinin-like immunoreactivity in central neurons innervating the salivary glands and hindgut of ixodid ticks. (2015)
  • Author(s): 59.Roller, L., Simo, L., Mizoguchi, A., Slovak M., Park, Y., Zitnan, D.
  • Journal Title: Cell Tiss. Res. Volume: 印刷中
  • Peer Reviewed
• [Presentation] ヨトウガ蛹休眠の休眠覚醒後の成虫発生を制御する内分泌機構 (2015)
  • Author(s): 山田信人、溝口明
  • Organizer: 日本動物学会第86回大会
  • Place of Presentation: 朱鷺メッセ（新潟）
  • Year and Date: 2015-09-17
• [Presentation] カイコガ生殖器原基の成長発達のホルモン調節機構 (2015)
  • Author(s): 藤永大輝、溝口明
  • Organizer: 日本動物学会第86回大会
  • Place of Presentation: 朱鷺メッセ（新潟）
  • Year and Date: 2015-09-17
• [Presentation] ヨトウガ蛹休眠の休眠覚醒を制御する内分泌機構 (2014)
  • Author(s): 山田信人、溝口明
  • Organizer: 日本動物学会第85回大会
  • Place of Presentation: 東北大学（仙台）
  • Year and Date: 2014-09-11
• [Presentation] カイコガ生殖器原基の成長発達のホルモン調節機構 (2014)
  • Author(s): 藤永大輝、河村祐亮、溝口明
  • Organizer: 日本動物学会第85回大会
  • Place of Presentation: 東北大学（仙台）
  • Year and Date: 2014-09-11
• [Book] Handbook of Hormones: Comparative Endocrinology for Basic and Clinical Research (2015)
  • Author(s): Mizoguchi, A.
  • Publisher: Elsevier