Selective labeling of target protein by using a chemical probe having a fluorogenic reactive group and an oligonucleotide aptamer

• Project/Area Number: 26810091
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Bio-related chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: Yamaguchi Takao 東京大学, 分子細胞生物学研究所, 助教 (80596601)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: アフィニティーラベル化 / 核酸アプタマー / タンパク質化学修飾 / 蛍光ラベル化

Outline of Final Research Achievements

Chemical probes, consist of an oligonucleotide aptamer and a fluorogenic reactive group, were designed to achieve a specific labeling of protein of interest (POI). Here, we prepared oligonucleotide aptamer for c-Met, a model POI. In addition, several fluorogenic reactive groups were designed and synthesized. Model experiments revealed that one of the designed fluorogenic reactive groups has a suitable stability and reactivity for the POI-specific labeling.

Research Products

• [Journal Article] Three-Dimensional QSAR Analysis and Design of New 1,2,4-Oxadiazole Antibiotics (2016)
  • Author(s): Erika Leemans, Kiran V. Mahasenan, Malika Kumarasiri, Edward Spink, Derong Ding, Peter I. O’Daniel, Marc A. Boudreau, Elena Lastochkin, Sebastian A. Testero, Takao Yamaguchi, Mijoon Lee, Dusan Hesek, Jed F. Fisher, Mayland Chang, Shahriar Mobashery
  • Journal Title: Bioorg. Med. Chem. Lett. Volume: 26 Issue: 3 Pages: 1011-1015
  • DOI: 10.1016/j.bmcl.2015.12.041
  • Peer Reviewed
• [Journal Article] Synthesis and Properties of 2’-O,4’-C-Spirocyclopropylene Bridged Nucleic Acid (scpBNA), an Analogue of 2’,4’-BNA/LNA Bearing a Cyclopropane Ring. (2015)
  • Author(s): Yamaguchi T., Horiba M., Obika S.
  • Journal Title: Chem. Commun. Volume: 23 Issue: 47 Pages: 33-37
  • DOI: 10.1039/c5cc02024g
  • Peer Reviewed
• [Journal Article] Synthesis and fluorescence properties of 4-diarylmethylene analogues of the green fluorescent protein chromophore (2015)
  • Author(s): Masahiro Ikejiri, Kousuke Matsumoto, Hiraku Hasegawa, Daisuke Yamaguchi, Moe Tsuchino, Yoshiko Chihara, Takao Yamaguchi, Kazuto Mori, Takeshi Imanishi, Satoshi Obika, Kazuyuki Miyashita
  • Journal Title: Tetrahedron Volume: 71 Issue: 30 Pages: 4987-4998
  • DOI: 10.1016/j.tet.2015.05.073
  • Peer Reviewed
• [Journal Article] Exploration of the Structure-Activity Relationship of 1,2,4-Oxadiazole Antibiotics (2015)
  • Author(s): Derong Ding, Marc A. Boudreau, Erika Leemans, Edward Spink, Takao Yamaguchi, Sebastian A. Testero, Peter I. O’Daniel, Elena Lastochkin, Mayland Chang, Shahriar Mobashery
  • Journal Title: Bioorg. Med. Chem. Lett. Volume: 25 Issue: 21 Pages: 4854-4857
  • DOI: 10.1016/j.bmcl.2015.06.044
  • Peer Reviewed
• [Journal Article] Discovery and structure-;activity relationship studies of N6-benzoyladenine derivatives as novel BRD4 inhibitors (2015)
  • Author(s): Noguchi-Yachide, T., Sakai, T., Hashimoto, Y., Yamaguchi, Y.
  • Journal Title: Bioorganic & Medicinal Chemistry Volume: 23 Issue: 5 Pages: 953-959
  • DOI: 10.1016/j.bmc.2015.01.022
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] アジドフタルイミドを発蛍光性光反応基とする新規アフィニティーラベル化法の開発 (2016)
  • Author(s): 千葉幸介、橋本祐一、山口卓男
  • Organizer: 日本薬学会 第136回年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-03-26
• [Presentation] 新規PPARαアゴニストの探索と構造展開、およびhACC2阻害活性評価 (2015)
  • Author(s): 岡崎翔吾、境太希、谷内出友美、石川稔、橋本祐一、山口卓男
  • Organizer: 第33回 メディシナルケミストリーシンポジウム
  • Place of Presentation: 幕張国際研修センター（千葉県・千葉市）
  • Year and Date: 2015-11-25
• [Presentation] Synthesis and properties of oligonucleotides containing 2’-O,4’-C-spirocyclopropylene bridged nucleic acids (scpBNA) having pyrimidine bases (2015)
  • Author(s): Masahiko Horiba, Takao Yamaguchi, Reiko Waki, Satoshi Obika
  • Organizer: SKO Symposium 2015
  • Place of Presentation: Seoul National University（韓国・ソウル市）
  • Year and Date: 2015-11-16
  • Int'l Joint Research
• [Presentation] Specific Fluorescence Labeling of a Target Protein Utilizing a Single, Small, Non-Fluorescent Unit (2015)
  • Author(s): Takao Yamaguchi, Kosuke Chiba, Yuichi Hashimoto, Kosuke Dodo, Miwako Asanuma, Mikiko Sodeoka
  • Organizer: Tokyo-LUM Symposium
  • Place of Presentation: Ludwig-Maximilians-Universitat（ドイツ・バイエルン州・ミュンヘン市）
  • Year and Date: 2015-10-28
  • Int'l Joint Research
• [Presentation] スピロシクロプロパン構造を導入した新規架橋型人工核酸の合成と物性評価 (2015)
  • Author(s): 堀場昌彦、山口卓男、小比賀聡
  • Organizer: 第41回 反応と合成の進歩シンポジウム
  • Place of Presentation: 近畿大学 11月ホール（大阪府・東大阪市）
  • Year and Date: 2015-10-26
• [Presentation] Synthesis and biophysical evaluation of 2’-O,4’-C-Spirocyclopropylene Bridged Nucleic Acids (scpBNA) (2014)
  • Author(s): Masahiko Horiba, Takao Yamaguchi, Satoshi Obika
  • Organizer: 第41回 国際核酸化学シンポジウム
  • Place of Presentation: 福岡、北九州国際会議場
  • Year and Date: 2014-11-05 – 2014-11-07
• [Presentation] 蛍光turn-ON型ラベル化法を用いた生物活性化合物の標的タンパク質の解析 (2014)
  • Author(s): 淺沼三和子、どど孝介、山口卓男、中西修一、斎藤洋平、岡﨑正晃、袖岡幹子
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都、国立京都国際会館
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] ニトロベンゾオキサジアゾールを用いる蛍光turn-ON型アフィニティーラベル化法の開発 (2014)
  • Author(s): 山口卓男、淺沼三和子、中西修一、斎藤洋平、岡﨑正晃、どど孝介、袖岡幹子
  • Organizer: 第12回 次世代を担う有機化学シンポジウム
  • Place of Presentation: 東京、日本薬学会長井記念ホール
  • Year and Date: 2014-05-23 – 2014-05-24
• [Remarks] 東京大学分子細胞生物学研究所 生体有機化学研究分野
  • URL: http://www.iam.u-tokyo.ac.jp/chem/IMCB-8ken-HP/Index.html
• [Patent(Industrial Property Rights)] 架橋型ヌクレオシドおよびヌクレオチド (2015)
  • Inventor(s): 小比賀聡、山口卓男、堀場昌彦、脇玲子
  • Industrial Property Rights Holder: 小比賀聡、山口卓男、堀場昌彦、脇玲子
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-02-17
  • Overseas