Possibility of skin care material using Pxt peptides from amphibian (Xenopus tropicalis) skin

• Project/Area Number: 15K01814
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biomolecular chemistry
• Research Institution: Wakayama Medical University (2018); National Institute of Advanced Industrial Science and Technology (2015-2017)
• Principal Investigator: Shigeri Yasushi 和歌山県立医科大学, 医学部, 教授 (90357187)
• Co-Investigator(Kenkyū-buntansha): 井村 知弘 国立研究開発法人産業技術総合研究所, 材料・化学領域, 研究グループ長 (10371022) ; 堀江 祐範 国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (30514591)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ペプチド / 表面張力 / ミセル / 溶血 / ナノディスク / 両生類 / 抗菌 / 立体構造解析 / 泡 / NMR / nanodisc / antimicrobial peptide / amphibian skin / surface activity / 炎症マーカー / ストレスマーカー

Outline of Final Research Achievements

Twelve novel peptides (Pxt-1-12) were isolated from the skin of Xenopus tropicalis, using topological mass spectrometry analysis. Several ion peaks corresponding to all identified Pxt peptides, were observed with MALDI-MS analysis of X. tropicalis secretory fluids, collected after in vivo stimulation suggesting that Pxt peptides were definitely secretory molecules. The remarkable foaming properties of Pxt-2 and Pxt-5 were observed. We further characterized Pxt-related peptides (Pxt-2, Pxt-5, Pxt-12, reverse Pxt-2, reverse Pxt-5, and reverse Pxt-12). All Pxt-related peptides formed significant amphiphilic alpha-helices. Moreover, Pxt-2, Pxt-5, and reverse Pxt-5 produced the micelle-like assemblies. Pxt-5 possessed the most potent hemolytic activity, while reverse Pxt-5 exhibited the most remarkable cytotoxicity in HaCaT cells. RT-PCR analysis confirmed the gene expressions of Pxt-5 in testis and Pxt-12 in muscle, in addition to skin, while Pxt-2 was only in skin.

Academic Significance and Societal Importance of the Research Achievements

カエル等の両生類は、皮膚を湿った状態に保ち、捕食者や微生物等の外敵から身を守るために、皮膚に多くの粘液腺を有し、抗菌や毒素を含んだ生体分子を積極的に分泌している。そこで独自に開発したトポロジカルマスを駆使した事により、両生類（ネッタイツメガエル）皮膚由来の新たな機能性生体分子（Pxtペプチド）を発見した。カタツムリの粘液由来のカタツムリクリーム、毒蛇由来の蛇毒クリーム（シンエイク, Syn-ake）なるものが最近の流行である事から、Pxtペプチドはこれまで知られていないコスメの添加成分として製品化研究に繋がり、コスメトロジーにおける新分野の開拓に貢献する可能性がある。

Research Products

• [Journal Article] Mass spectrometry analysis and biological characterization of the venom and venom sac components of the predatory ant Odontomachus monticola. (2019)
  • Author(s): Tani, N., Kazuma, K., Ohtsuka, Y., Shigeri, Y., Masuko, K., Konno, K., Inagaki, H.
  • Journal Title: Toxins Volume: 10 Issue: 1 Pages: 50-50
  • DOI: 10.3390/toxins11010050
  • Peer Reviewed / Open Access
• [Journal Article] Lipid Nanodisc Formation using Pxt-5 Peptide Isolated from Amphibian (<i>Xenopus tropicalis</i>) Skin, and its Altered Form, Modify-Pxt-5 (2018)
  • Author(s): Ikeda, Y., Taira, T., Sakai, K., Sakai, H., Shigeri, Y., Imura, T.
  • Journal Title: Journal of Oleo Science Volume: 67 Issue: 8 Pages: 1035-1041
  • DOI: 10.5650/jos.ess18051
  • NAID: 130007429988
  • ISSN: 1345-8957, 1347-3352
  • Peer Reviewed / Open Access
• [Journal Article] Modification of N-terminal amino acids of fungal benzoate hydroxylase (CYP53A15) for the production of p-hydroxybenzoate and optimization of bioproduction conditions in Escherichia coli. (2018)
  • Author(s): Tamaki, S., Yagi, M., Nishihata, Y., Yamaji, H., Shigeri, Y., Uno, T., and Imaishi, H.
  • Journal Title: J. Microbiol. Biotech. Volume: 28 Issue: 3 Pages: 439-447
  • DOI: 10.4014/jmb.1711.11030
  • Peer Reviewed
• [Journal Article] Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata. (2017)
  • Author(s): Kawakami, H., Goto, S., Murata, K., Matsuda, H., Shigeri, Y., Imura, T., Inagaki, H., and Shinada, T.
  • Journal Title: Journal of Venomous Animals and Toxins including Tropical Diseases Volume: 23 Issue: 1 Pages: 1-11
  • DOI: 10.1186/s40409-017-0119-6
  • Peer Reviewed / Open Access
• [Journal Article] Novel biologically active peptides isolated from the venom of Xylocopa appendiculata. (2017)
  • Author(s): Kawakami, H., Goto, S., Murata, K., Matsuda, H., Shigeri, Y., Imura, T., Inagaki, H., and Shinada, T.
  • Journal Title: Journal of Venomous Animals and Toxins including Tropical Diseases Volume: 印刷中
  • Peer Reviewed
• [Journal Article] Physicochemical and biological characterizations of Pxt peptides from amphibian (Xenopus tropicalis) skin. (2016)
  • Author(s): Shigeri, Y., Horie, M., Yoshida, T., Hagihara, Y., Imura, T., Inagaki, H., Haramoto, Y., Ito, Y., and Asashima, M.
  • Journal Title: J. Biochem. Volume: 159 Issue: 6 Pages: 619-629
  • DOI: 10.1093/jb/mvw003
  • NAID: 40020869436
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Physicochemical and biological characterizations of Pxt peptides from amphibian (Xenopus tropicalis) skin (2016)
  • Author(s): Yasushi Shigeri, Masanori Horie, Tsuyoshi Yoshida, Yoshihisa Hagihara, Tomohiro Imura, Hidetoshi Inagaki, Yoshikazu Haramoto, Yuzuru Ito, and Makoto Asashima
  • Journal Title: Journal of Biochemistry Volume: -
  • NAID: 40020869436
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] アギトアリ毒液成分のペプチドーム及びプロテオーム解析 (2019)
  • Author(s): 谷直紀、数馬恒平、大塚幸雄、茂里康、増子恵一、紺野勝弘、稲垣英利
  • Organizer: 第三回京都生体質量分析研究会シンポジウム
• [Presentation] 熱帯ツメガエルの分泌ペプチドを利用した脂質ナノディスクの調製 (2017)
  • Author(s): 池田ゆり、茂里康、井村知弘
  • Organizer: 日本化学会第97春季年会
  • Place of Presentation: 神奈川県横浜市
  • Year and Date: 2017-03-17
• [Presentation] 熱帯ツメガエルの分泌ペプチドを利用した脂質ナノディスクの調製。 (2017)
  • Author(s): 池田ゆり、茂里康、井村知弘
  • Organizer: 日本化学会第97春季年会(2017)
• [Presentation] 両生類（ネッタイツメガエル）皮膚由来、機能性ペプチドの網羅的探索 (2015)
  • Author(s): 茂里康
  • Organizer: 第32回関西地区ペプチドセミナー
  • Place of Presentation: 毎日インテシオ（大阪府）
  • Year and Date: 2015-11-28
  • Invited
• [Patent(Industrial Property Rights)] カルモジュリン依存性タンパク質リン酸化酵素Ｉ由来ポリペプチド、リン酸化剤、および製造方法 (2016)
  • Inventor(s): 千賀由佳子、秋月一駿、片山将一、茂里康 et al
  • Industrial Property Rights Holder: 千賀由佳子、秋月一駿、片山将一、茂里康 et al
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2016-095770
  • Filing Date: 2016-05-12
• [Patent(Industrial Property Rights)] 両生類ネッタイツメガエル皮膚由来機能性ペプチド (2015)
  • Inventor(s): 茂里康、萩原義久、渡部邦彦、井村知弘、原本悦和、伊藤弓弦、浅島誠
  • Industrial Property Rights Holder: 茂里康、萩原義久、渡部邦彦、井村知弘、原本悦和、伊藤弓弦、浅島誠
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015
  • Acquisition Date: 2018
• [Patent(Industrial Property Rights)] 界面活性ペプチドから短時間で形成する巨大ナノディスク (2015)
  • Inventor(s): 井村知弘、茂里康、遠藤明、平敏彰
  • Industrial Property Rights Holder: 井村知弘、茂里康、遠藤明、平敏彰
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-163190
  • Filing Date: 2015-08-20