Highly efficient cell-free membrane protein expression in organic-inorganic hybrid vesicles

• Project/Area Number: 24681028
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Microdevices/Nanodevices
• Research Institution: Nara Institute of Science and Technology
• Principal Investigator: Yasuhara Kazuma 奈良先端科学技術大学院大学, 物質創成科学研究科, 助教 (90545716)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥27,300,000 (Direct Cost: ¥21,000,000、Indirect Cost: ¥6,300,000); Fiscal Year 2015: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2014: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2013: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2012: ¥16,380,000 (Direct Cost: ¥12,600,000、Indirect Cost: ¥3,780,000)
• Keywords: バイオリアクター / マイクロ・ナノデバイス / 有機－無機ハイブリッド / ベシクル / 遺伝子発現 / 有機ー無機ハイブリッド / 膜タンパク質 / タンパク質 / 有機-無機ハイブリッド / 蛋白質

Outline of Final Research Achievements

This study focused on the development of a highly efficient cell-free membrane protein expression system using organic-inorganic hybrid vesicles. The organic-inorganic hybrid vesicles with a crosslinked ceramic-like surface have a semi-permeable feature in which small molecules can permeate across the membrane whereas large molecules cannot. The structure and semi-permeable feature of the organic-inorganic hybrid vesicles can be tuned by the incorporation of various amphiphiles. It was suggested that the encapsulation of enzymatic reaction system enables the development of a membrane protein expression system, which utilizes the semi-permeable feature of the organic-inorganic hybrid vesicles.

Research Products

• [Int'l Joint Research] ミシガン大学(米国)
• [Journal Article] Anisotropic Self-Assembly of Citrate-Coated Gold Nanoparticles on Fluidic Liposomes (2016)
  • Author(s): K. Sugikawa, T. Kadota, K. Yasuhara, A. Ikeda
  • Journal Title: Angew. Chem. Int. Ed. Volume: 55 Issue: 12 Pages: 4059-4063
  • DOI: 10.1002/anie.201511785
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Kinetic study of all-or-none hemolysis induced by catonic amphiphilic polymethacrylates with antimicorbial activity (2015)
  • Author(s): Kazuma Yasuhara, Kenichi Kuroda
  • Journal Title: Chinese Chemical Letters Volume: 印刷中 Issue: 4 Pages: 479-484
  • DOI: 10.1016/j.cclet.2015.01.029
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Lipid-Membrane-Incorporated Hydrophobic Photochromic Molecules Prepared by the Exchange Method Using Cyclodextrins (2015)
  • Author(s): A. Ikeda, S. Hino, K. Ashizawa, K. Sugikawa, J. Kikuchi, M. Tsukamoto, K. Yasuhara
  • Journal Title: Org. Biomol. Chem. Volume: 13 Issue: 22 Pages: 0-0
  • DOI: 10.1039/c5ob00240k
  • NAID: 120006469197
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Porphyrin-uptake in liposomes and living cells using an exchange method with cyclodextrin (2015)
  • Author(s): A. Ikeda, S. Hino, T. Mae, Y. Tsuchiya, K. Sugikawa, M. Tsukamoto, K. Yasuhara, H. Shigeto, H. Funabashi, A. Kuroda, M. Akiyama
  • Journal Title: RSV Adv. Volume: 5 Issue: 127 Pages: 105279-105287
  • DOI: 10.1039/c5ra24985f
  • Peer Reviewed / Open Access
• [Journal Article] [70]Fullerenes Assists the Formation of Phospholipid Bicelles at Low Lipid Concentrations. (2014)
  • Author(s): A. Ikeda, K. Kiguchi, T. Hida, K. Yasuhara, K. Nobusawa, M. Akiyama, W. Shinoda
  • Journal Title: Langmuir Volume: 30 Issue: 41 Pages: 12315-12320
  • DOI: 10.1021/la503732q
  • NAID: 120006469202
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Dynamic behaviour in giant unilamellar vesicles induced by the uptake of [70]fullerene (2014)
  • Author(s): A. Ikeda, T. Hida, T. Iizuka, M. Tsukamoto, J. Kikuchi and K. Yasuhara
  • Journal Title: Chem. Commun. Volume: 50 Issue: 11 Pages: 1288-1291
  • DOI: 10.1039/c3cc47711h
  • Peer Reviewed
• [Journal Article] (2014)
  • Author(s): S. Junedi, K. Yasuhara, S. Nagao, J. Kikuchi, S. Hirota
  • Journal Title: ChemBioChem Volume: Vol. 15, No. 4 Issue: 4 Pages: 517-521
  • DOI: 10.1002/cbic.201300728
  • Peer Reviewed
• [Journal Article] Modulation of raft domains in a lipid bilayer by boundary-active curcumin (2014)
  • Author(s): M. Tsukamoto, K. Kuroda, A. Ramamoorthy and K. Yasuhara
  • Journal Title: Chem. Commun. Volume: 50 Issue: 26 Pages: 3427-3430
  • DOI: 10.1039/c3cc47738j
  • Peer Reviewed
• [Journal Article] Synthetic cell division system: Controlling equal vs. unequal divisions by design (2013)
  • Author(s): Yoichi Sato, et. al.
  • Journal Title: Scientific Reports Volume: 3 Issue: 1 Pages: 3475-3475
  • DOI: 10.1038/srep03475
  • Peer Reviewed
• [Journal Article] Spontaneously Formed Semipermeable Organic-Inorganic Hybrid Vesicles Permitting Molecular Weight Selective Transmembrane Passage (2013)
  • Author(s): K. Yasuhara, T. Kawasaki, S. Okuda, S. Oshima, J. Kikuchi
  • Journal Title: Chem. Commun Volume: 49(7) Issue: 7 Pages: 665-667
  • DOI: 10.1039/c2cc36662b
  • Peer Reviewed
• [Journal Article] セラミック表面によって安定化された脂質二分子膜ナノディスクの形成 (2013)
  • Author(s): 安原主馬
  • Journal Title: C&I commun Volume: 38 Pages: 29-32
  • NAID: 40020187323
• [Journal Article] Thermal Stability of Synthetic Lipid Bicelles Encompassed by Siloxane Surfaces as Organic-Inorganic Hybrid Nanodiscs (2012)
  • Author(s): K. Yasuhara, H. Hayashi J. Kikuchi
  • Journal Title: Chem. Lett Volume: 41(10) Issue: 10 Pages: 1223-1225
  • DOI: 10.1246/cl.2012.1223
  • NAID: 130004426360
  • Peer Reviewed
• [Presentation] 両親媒性高分子による脂質膜の断片化とナノディスク形成 (2016)
  • Author(s): 荒木田臣・菊池純一・安原主馬
  • Organizer: 日本化学会第96春季年会
  • Place of Presentation: 同志社大学 京田辺キャンパス（京都府京田辺市）
  • Year and Date: 2016-03-24
• [Presentation] Design of Lipid Molecules for Specific Domain Formation in Artificial Cell Membranes (2016)
  • Author(s): Yuri, Ishikura, Yoshiyuki Horiuchi, Taku Yamazaki,Kazuma Yasuhara, Jun-ichi Kikuchi
  • Organizer: 日本化学会第96春季年会
  • Place of Presentation: 同志社大学 京田辺キャンパス（京都府京田辺市）
  • Year and Date: 2016-03-24
• [Presentation] SPONTANEOUS LIPID NANODISC FORMATION BY MEMBRANE-ACTIVE POLYMERS (2016)
  • Author(s): K. Yasuhara, J. Arakida, J. Kikuchi
  • Organizer: 60th Biophysical Society Annual Meeting
  • Place of Presentation: Los Angeles Convention Center, Los Angeles, CA, USA
  • Year and Date: 2016-02-27
  • Int'l Joint Research
• [Presentation] Designed amphiphilic polymers for lipid membrane nanodisc formation (2015)
  • Author(s): K. Yasuhara, J. Arakida, J. Kikuchi
  • Organizer: Pacifichem 2015
  • Place of Presentation: Hawaii Convention Center, Honolulu, HI, USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research
• [Presentation] Design and evaluation of membrane-active polymers to form lipid bilayer nanodisc (2015)
  • Author(s): K. Yasuhara, J. Arakida, J. Kikuchi
  • Organizer: Macromolecular Science & Engineering 39th Annual Symposium
  • Place of Presentation: University of Michigan, Ann Arbor, MI, USA
  • Year and Date: 2015-10-22
  • Int'l Joint Research
• [Presentation] 脂質ナノディスクを形成する膜活性ポリマーの設計 (2015)
  • Author(s): 荒木田臣・菊池純一・安原主馬
  • Organizer: 第64回高分子討論会
  • Place of Presentation: 東北大学 川内キャンパス（宮城県仙台市）
  • Year and Date: 2015-09-15
• [Presentation] Membrane-active amphiphilic polymers for lipid nanodisc formaiton (2015)
  • Author(s): 安原主馬・荒木田臣・菊池純一
  • Organizer: 第53回生物物理学会年会
  • Place of Presentation: 金沢大学 角間キャンパス（石川県金沢市）
  • Year and Date: 2015-09-13
• [Presentation] 脂質ナノディスクを形成する膜活性ポリマー (2015)
  • Author(s): 安原主馬・荒木田臣・菊池純一
  • Organizer: 第66回コロイドおよび界面化学討論会
  • Place of Presentation: 鹿児島大学 郡元キャンパス（鹿児島県鹿児島市）
  • Year and Date: 2015-09-10
• [Presentation] Membrane-active amphiphilic polymers for lipid nanodisc formation (2015)
  • Author(s): K. Yasuhara, J. Arakida, J. Kikuchi
  • Organizer: IACIS 2015
  • Place of Presentation: Convention Center Rheingoldhalle, Mainz, Germany
  • Year and Date: 2015-05-24
  • Int'l Joint Research
• [Presentation] Self-Assembly of Organic-Inorganic Hybrid Lipids in Water Toward Fabrication of Bio-nanomaterials (2014)
  • Author(s): K. Yasuhara
  • Organizer: RPI MTLE department seminar
  • Place of Presentation: Troy, NY / USA
  • Year and Date: 2014-11-13
  • Invited
• [Presentation] Fablication of semipermeable organic-inorganic hybrid vesicles and their application to nanocapsule for enzymatic reaction (2014)
  • Author(s): K. Yasuhara, J. Kikuchi
  • Organizer: "The 7th International Workshop on ADVANCED MATERIALS SCIENCE AND NANOTECHNOLOGY "
  • Place of Presentation: Ha Long City / Vietnam
  • Year and Date: 2014-11-02 – 2014-11-06
  • Invited
• [Presentation] Interaction of Zinc-Chelating Polymer with Cell Membranes (2014)
  • Author(s): M. Namba, M. Tsukamoto, J. Kikuchi, K. Yasuhara
  • Organizer: Macromolecular Science & Engineering 38th Annual Symposium
  • Place of Presentation: Ann Arbor, MI / USA
  • Year and Date: 2014-10-23
• [Presentation] Organic-inorganic Hybrid Vesicles as Spontaneously Formed Semipermeable Nanocapsule (2014)
  • Author(s): K. Yasuhara, J. Kikuchi
  • Organizer: XXIII INTERNATIONAL MATERIALS RESEARCH CONGRESS 2014
  • Place of Presentation: Cancun / Mexico
  • Year and Date: 2014-08-17 – 2014-08-21
• [Presentation] 有機無機ハイブリッドベシクルの選択的膜透過を利用した酵素反応制御 (2013)
  • Author(s): 大島匠平・安原主馬・菊池純一
  • Organizer: 日本化学会第93春季年会
  • Place of Presentation: 立命館大学(滋賀県)(ポスター発表)
  • Year and Date: 2013-03-24
• [Presentation] 有機-無機ハイブリッドベシクルの分子ふるい機能とバイオナノリアクターへの応用 (2013)
  • Author(s): 安原主馬・大島匠平・菊池純一
  • Organizer: 日本化学会第93春季年会
  • Place of Presentation: 立命館大学(滋賀県)
  • Year and Date: 2013-03-23
• [Presentation] カチオン性カリックスアレーン誘導体の抗菌活性 (2013)
  • Author(s): 永井佐知・安原主馬・菊池純一
  • Organizer: 第62回高分子学会年次大会
  • Place of Presentation: 京都国際会館（京都府京都市)
• [Presentation] 抗菌性カリックスアレーン誘導体の構造-活性相関 (2013)
  • Author(s): 永井佐知・安原主馬・菊池純一
  • Organizer: 第7回バイオ関連シンポジウム
  • Place of Presentation: 名古屋大学（愛知県名古屋市）
• [Presentation] Supontaneously formed semipermeable organic-inorganic hybrid vesicles and their application to bio-nano reactors (2013)
  • Author(s): K. Yasuhara, J. Kikuchi
  • Organizer: XVII International Sol-Gel Conference
  • Place of Presentation: Melia Castilla Hotel (Madrid, Spain)
• [Presentation] Organic-Inorganic Hybrid Vesicle as Semipermeable Nano-capsule (2012)
  • Author(s): K. Yasuhara, J. Kikuchi
  • Organizer: The Macro/NAIST Joint Workshop for Emerging Researchers
  • Place of Presentation: University of Michigan Ann Arbor, MI USA(招待講演)
  • Year and Date: 2012-10-24
• [Presentation] 有機-無機ハイブリッドベシクル「セラソーム」における分子量選択的膜透過とその制御 (2012)
  • Author(s): 安原主馬, 奥田静代, 菊池純一
  • Organizer: 日本ゾル-ゲル学会 第10回討論会
  • Place of Presentation: 慶應義塾大学(東京都)(ポスター発表)
  • Year and Date: 2012-07-27
• [Presentation] セラソームの半透膜機能とその制御 (2012)
  • Author(s): 安原主馬, 奥田静代, 大島匠平, 菊池純一
  • Organizer: 第61回高分子学会年次大会
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2012-05-29
• [Presentation] 有機-無機ハイブリッド脂質の自己組織化とバイオ応用
  • Author(s): 安原主馬
  • Organizer: 奈良先端未来開拓コロキウム
  • Place of Presentation: 奈良先端科学技術大学院大学（奈良県生駒市）
• [Presentation] Antimicrobial activity of amphiphilic calix[4]arene derivatives
  • Author(s): S. Nagai, K. Yasuhara, J. Kikuchi
  • Organizer: Macromolecular Science & Engineering 37th Annual Symposium
  • Place of Presentation: University of Michigan (Ann Arbor, MI USA)