Development of chemical methods for analyzing protein molecules function in their natural habitat

2013 Fiscal Year Final Research Report

• Project Area: Integrative understanding of biological processes mediated by transient macromolecular complexes; New technology for visualizing physiologically metastable states.
• Project/Area Number: 21121005
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: Kyoto University
• Principal Investigator: HAMACHI Itaru 京都大学, 工学(系)研究科(研究院), 教授 (90202259)
• Co-Investigator(Kenkyū-buntansha): OJIDA Akio 九州大学, 大学院薬学研究院, 教授 (10343328)
• Project Period (FY): 2009-07-23 – 2014-03-31
• Keywords: タンパク質 / 小分子プローブ / ラベル化 / 有機化学 / イメージング

Research Abstract

Protein is one of the central biomolecules that is involved in almost all biological events and thus its quantitative analysis has been actively conducted for many years. Recently, it is now regarded that protein analysis in natural habitats (living systems is ideal) should be crucial, in addition to the analysis in the purified test tube systems. In this research project, we successfully developed several original methods (or synthetic small molecules) based on chemistry that allow for selective labeling and imaging target proteins under nearly natural conditions. These methods may contribute to analyzing protein structure and functions in living cells.

Research Products

• [Journal Article] Synthetic Self-Localizing Ligands That Control The Spatial Location of Proteins in Living Cells (2013)
  • Author(s): M. Ishida, H. Watanabe, K. Takigawa, Y. Kurishita, C. Oki, A. Nakamura, I. Hamachi, S. Tsukiji
  • Journal Title: J. Am. Chem. Soc Volume: 135 Pages: 12684-12689
  • DOI: 10.1021/ja4046907
  • Peer Reviewed
• [Journal Article] Semisynthetic Lectin-Dimethylaminopyridine Conjugates for Labeling and Profiling Glycoproteins on Live Cell Surfaces (2013)
  • Author(s): T. Hayashi, Y. Sun, T. Tamura, K. Kuwata, Z. Song, Y. Takaoka, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 135 Pages: 12252-12258
  • DOI: 10.1021/ja4043214
  • Peer Reviewed
• [Journal Article] Fluorophore labeling of native FKBP12 by ligand-directed tosyl chemistry allows detection of its molecular interactions in vitro and in living cells (2013)
  • Author(s): T. Tamura, Y. Kioi, T. Miki, S. Tsukiji, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 135 Pages: 6782-6785
  • DOI: 10.1021/ja401956b
  • Peer Reviewed
• [Journal Article] One-step construction of caged carbonic anhydrase I using a ligand-directed acyl imidazole-based protein labeling method (2013)
  • Author(s): K. Matsuo, Y. Kioi, R. Yasui, Y. Takaoka, T. Miki, S. Fujishima, I. Hamachi
  • Journal Title: Chemical Science Volume: 4 Pages: 2573-2580
  • DOI: 10.1039/c3sc50560j
  • Peer Reviewed
• [Journal Article] Organelle-Localizable Fluorescent Chemosensors for Site-Specific Multicolor Imaging of Nucleoside Polyphosphate Dynamics in Living Cells (2012)
  • Author(s): Y. Kurishita, T. Kohira, A. Ojida, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 134 Pages: 18779-18789
  • DOI: 10.1021/ja308754g
  • Peer Reviewed
• [Journal Article] Ligand-directed Acyl Imidazole Chemistry for Labeling of Membrane-bound Proteins on Live Cells (2012)
  • Author(s): S. Fujishima, R. Yasui, T. Miki, A. Ojida, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 134 Pages: 3961-3964
  • DOI: 10.1021/ja2108855
  • Peer Reviewed
• [Journal Article] Native FKBP12 Engineering by Ligand-Directed Tosyl Chemistry : Labeling Properties and Application to Photo-Cross-Linking of Protein Complexes in Vitro and in Living Cells (2012)
  • Author(s): T. Tamura, S. Tsukiji, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 134 Pages: 2216-2226
  • DOI: 10.1021/ja209641t
  • Peer Reviewed
• [Journal Article] Chemical Cell-Surface Receptor Engineering Using Affinity-Guided, Multivalent Organocatalysts (2011)
  • Author(s): H. Wang, Y. Koshi, D. Minato, H. Nonaka, S. Kiyonaka, Y. Mori, S. Tsukiji, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 133 Pages: 12220-12228
  • DOI: 10.1021/ja204422r
  • Peer Reviewed
• [Journal Article] Selective Covalent Labeling of Tag-Fused GPCR Proteins on Live Cell Surface with a Synthetic Probe for Their Functional Analysis (2010)
  • Author(s): H. Nonaka, S. Fujishima, S. Uchinomiya, A. Ojida, I. Hamachi
  • Journal Title: J. Am. Chem. Soc Volume: 132 Pages: 9301-9309
  • DOI: 10.1021/ja910703v
  • Peer Reviewed
• [Journal Article] Self-assembling nano-probes displaying off/on 19F NMR signals for protein detection and imaging (2009)
  • Author(s): Y. Takaoka, T. Sakamoto, S. Tsukiji, M. Narazaki, T. Matsuda, H. Tochio, M. Shirakawa, I. Hamachi
  • Journal Title: Nature Chemistry Volume: 1 Pages: 557-561
  • DOI: 10.1038/NCHEM.365
  • Peer Reviewed
• [Presentation] Selective Chemical Labeling and Imaging of Endogenous Protein in Live Cells (2013)
  • Author(s): Itaru Hamachi
  • Organizer: 23rd American Peptide Symposium
  • Place of Presentation: Hilton Waikoloa Village, Hawaii
  • Year and Date: 20130622-27
• [Presentation] 蛋白質ケミカルラべリングを基盤とした細胞有機化学 (2013)
  • Author(s): 浜地 格
  • Organizer: 第13回日本蛋白質科学会年会
  • Place of Presentation: とりぎん文化会館
  • Year and Date: 20130612-14
• [Presentation] 細胞夾雑系でのタンパク質合成化学 (2013)
  • Author(s): 浜地 格
  • Organizer: 日本化学会第93春季年会
  • Place of Presentation: 立命館大学びわこ・くさつキャンパス
  • Year and Date: 20130322-25
• [Presentation] 生細胞での化学プローブ/蛋白質の直接ハイブリッド化による機能化 (2013)
  • Author(s): 浜地 格
  • Organizer: 化学工学会第45回秋季大会
  • Place of Presentation: 岡山大学
  • Year and Date: 2013-09-16
• [Presentation] 内在性蛋白質のケミカルラベル化法の開発とイメージング (2013)
  • Author(s): 浜地 格
  • Organizer: 第22回日本バイオイメージング学会学術集会
  • Place of Presentation: 東京大学
  • Year and Date: 2013-09-15
• [Presentation] 蛋白質ケミカルラベリングを基軸とした細胞有機化学 (2012)
  • Author(s): 浜地 格
  • Organizer: 第30回メディシナルケミストリーシンポジウム
  • Place of Presentation: タワーホール船堀
  • Year and Date: 20121128-30
• [Presentation] タンパク質ラベリングを基盤とする生細胞化学 (2012)
  • Author(s): 浜地 格
  • Organizer: 日本化学会第92春季年会
  • Place of Presentation: 慶應義塾大学日吉キャンパス・矢上キャンパス
  • Year and Date: 20120325-28
• [Presentation] Protein labeling and engineering in live cell systems (2011)
  • Author(s): Itaru Hamachi
  • Organizer: CIPSM -Fest of Chemical Biology
  • Place of Presentation: Buchner Lecture Hall, Ludwig Maximilians University Munich
  • Year and Date: 20110915-16
• [Presentation] Chemical Labeling and Imaging of Protein in Live CellsI (2011)
  • Author(s): Itaru Hamachi
  • Organizer: UPAC International Congress on Analytical Science 2011
  • Place of Presentation: Kyoto International Conference Center
  • Year and Date: 20110523-25
• [Presentation] A Chemistry-Based Method for Labeling membrane- Bound Proteins on Live Cell Surface (2011)
  • Author(s): Itaru Hamachi
  • Organizer: The 5th International Workshop on Approaches to Single-Cell Analysis
  • Place of Presentation: The Takeda Hall,The University of Tokyo
  • Year and Date: 20110303-04
• [Presentation] Chemistry-based protein labeling and imaging in cell and in vivo (2010)
  • Author(s): Itaru Hamachi
  • Organizer: 5th International Peptide Symposium
  • Place of Presentation: Kyoto International Conference Center
  • Year and Date: 20101204-09
• [Presentation] Chemistry-Based Methods for Selective Lectin Labeling (2010)
  • Author(s): Itaru Hamachi
  • Organizer: ICS2010
  • Place of Presentation: Makuhari Messe International Convention Complex
  • Year and Date: 20100801-06
• [Presentation] 内在性蛋白質の選択的ラベル化とイメージング (2010)
  • Author(s): 浜地 格
  • Organizer: 日本化学会第90春季年会
  • Place of Presentation: 近畿大学
  • Year and Date: 20100326-29
• [Remarks]
  • URL: http://www.sbchem.kyoto-u.ac.jp/hamachi-lab