外来物質との相互作用や外場による生体膜のトポロジー変化のダイナミクスとメカニズム

• Project Area: Creation of non-equilibrium soft matter physics: Structure and dynamics of mesoscopic systems
• Project/Area Number: 21015009
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Shizuoka University
• Principal Investigator: 山崎 昌一 静岡大学, 創造科学技術大学院, 教授 (70200665)
• Co-Investigator(Kenkyū-buntansha): 岡 俊彦 静岡大学, 理学部, 講師 (60344389)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥7,800,000 (Direct Cost: ¥7,800,000); Fiscal Year 2010: ¥3,900,000 (Direct Cost: ¥3,900,000); Fiscal Year 2009: ¥3,900,000 (Direct Cost: ¥3,900,000)
• Keywords: 生物物理学 / ナノバイオ / 生体膜 / キュービック相 / 2分子膜液晶相 / 相転移 / 静電相互作用 / 時分割X線小角散乱法

Research Abstract

我々は生体膜の2分子膜液晶相(L_α相)とキュービック相(Q相)の間の相転移が静電相互作用により起こることを初めて見出し、その後系統的な研究を進めてきた。最近、中性でL_α相を形成する20%ジオレオイルホスファチジルセリン(DOPS)と80%モノオレイン(MO)の混合膜の多重層リポソーム(20%DOPS/80%MO-MLV)の水溶液のpHを下げていくと、最終pHが2.9以下のときにL_α相からQ_<11>^D相(Q^<244>相)への相転移が1時間以内に起こることを発見した。この相転移のメカニズムやキネティックスパスウエイを解明するために、SPring-8やPFの放射光を用いて、膜の構造変化を時分割X線小角散乱(TR-SAXS)により研究した。
まず、中性の緩衝液中で作成した20%DOPS/80%MO-MLVの懸濁液と9倍量の低いpHの緩衝液を自作の二液混合装置を用いて急速に混合し、その後の膜の構造変化をTR-SAXSにより測定した。5%(w/v)のポリエチレングリコール6000(PEG-6K)存在下で最終pHが2.6から2.9の時は、混合後2-10秒以内にL_α相のピークが消失するとともにヘキサゴナルII(H_<11>)相のピークが成長し、30-90s後からQ <II>^D相のピークが現れ始め、その後ゆっくりとH_<II>相からQ_<II>^D相への相転移が起こり、15-30分以内でH_<II>相のピークが消失した。この実験結果から、低いpHが誘起するL_α相からQ_<II>^D相への相転移では、まずL_α相から急速にH_<II>相に相転移し、その後ゆっくりQ_<II>^D相へ相転移することがわかった。さらにこの相転移の後半の過程であるH_<II>相からQ_<II>^D相への相転移の速度定数を特異値分解法により求めた。これらの結果に基づいて、低いpHが誘起するL_α相からQ_<II>^D相への相転移のメカニズムを考察した。

Research Products

• [Journal Article] Spontaneous insertion of lipopolysaccharide into lipid membranes from aqueous solution (2011)
  • Author(s): J.M.Alam
  • Journal Title: Chem.Phys.Lipids Volume: 164 Pages: 166-174
  • Peer Reviewed
• [Journal Article] Kinetics of Low pH-Induced Lamellar to Bicontinuous Cubic Phase Transition in Dioleoylphosphatidylserine/Monoolein (2011)
  • Author(s): M.M.Alam
  • Journal Title: J.Chem.Phys. Volume: 134
  • Peer Reviewed
• [Journal Article] A membrane filtering method for the purification of giant unilamellar vesicles (2011)
  • Author(s): Y.Tamba
  • Journal Title: Chem.Phys.Lipids Volume: 164(In press)
  • Peer Reviewed
• [Journal Article] 膜で測る:生体膜の機能やダイナミクスを解明する単一GUV法 (2011)
  • Author(s): 山崎昌一
  • Journal Title: 実験医学 Volume: 29(In press)
• [Journal Article] Kinetic pathway of antimicrobial peptide magainin 2-induced pore formation in lipid membranes (2010)
  • Author(s): Y.Tamba
  • Journal Title: J.Phys.Chem.B Volume: 114 Pages: 12018-12026
  • Peer Reviewed
• [Journal Article] 単一GUV法による抗菌ペプチドのポア形成の研究 (2010)
  • Author(s): 山崎昌一
  • Journal Title: 生物物理 Volume: 50 Pages: 296-297
  • NAID: 10026965330
  • Peer Reviewed
• [Journal Article] Transformation between Liposomes and Cubic Phases of Biological Lipid Membranes Induced by Modulation of Electrostatic Interactions (2009)
  • Author(s): M.Yamazaki
  • Journal Title: Advances in Planar Lipid Bilayers and Liposomes 9 Pages: 163-209
• [Journal Article] 単一GUV法を用いたペプヂドと脂質膜の相互作用の解析 (2009)
  • Author(s): 山崎昌一
  • Journal Title: 膜 34 Pages: 126-132
  • Peer Reviewed
• [Journal Article] The Size of the Pore in Lipid Membranes Induced by Antimicrobial Peptide Magainin 2 (2009)
  • Author(s): H.Ariyama
  • Journal Title: 2009 IEEE International Symposium on Micro-NanoMechatronics and Human Science Pages: 208-213
• [Journal Article] Magainin 2-induced pore formation in membrane depends on its concentration in membrane interface (2009)
  • Author(s): Y.Tamba
  • Journal Title: J.Phys.Chem.B 113 Pages: 4846-4852
  • Peer Reviewed
• [Presentation] 外来物質との相互作用による生体膜のトポロジー変化のダイナミクスとメカニズム (2011)
  • Author(s): 山崎昌一
  • Organizer: 特定領域研究「ソフトマター物理」第5回領域研究会
  • Place of Presentation: 東京(東京大学・弥生講堂)
  • Year and Date: 2011-01-07
• [Presentation] The Single GUV Method Reveals the Antimicrobial Peptide-Induced Pore Formation in Lipid Membranes (2010)
  • Author(s): M.Yamazaki
  • Organizer: BIT's 1st Annual World Congress of NanoMedicine-2010
  • Place of Presentation: Beijing International Convention Center, Beijing, China(招待講演)
  • Year and Date: 2010-10-25
• [Presentation] Effect of transportan-10 on membrane permeability of single DOPC-GUVs (2010)
  • Author(s): H.Ariyama
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-22
• [Presentation] Membrane filtering method for purification of giant unilamellar vesicles (2010)
  • Author(s): Y.Tamba
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-22
• [Presentation] Construction of microarray of GUVs of oil-free membranes for single GUV method (2010)
  • Author(s): H.Terashima
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-22
• [Presentation] Protein toxin lysenin-induced pore formation in lipid membranes : the single GUV method study (2010)
  • Author(s): R.Takagi
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-21
• [Presentation] Mechanism of subcritical pore formation in charged lipid membranes induced by antimicrobial peptide-magainin 2 (2010)
  • Author(s): V.Levadny
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-21
• [Presentation] Effect of mechanical tension on the leakage of internal contents of liposomes of biomembranes (2010)
  • Author(s): T.Takahashi
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-21
• [Presentation] Kinetic pathway of antimicrobial peptide magainin 2-induced pore formation in lipid membranes (2010)
  • Author(s): M.Yamazaki
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-21
• [Presentation] Lipopolysaccharide-induced shape changes and vesicle fission of giant unilamellar vesicles of lipid membranes (2010)
  • Author(s): J.M.Alam
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-20
• [Presentation] Kinetic of low-pH-induced transformation of bilayer membrane to cubic phases in DOPS/MO membranes (2010)
  • Author(s): M.M.Alam
  • Organizer: 日本生物物理学会第48回年次大会
  • Place of Presentation: 仙台(東北大学・川内北キャンパス)
  • Year and Date: 2010-09-20
• [Presentation] Kinetic Pathway of Antimicrobial Peptide Magainin 2-Induced Pore Formation in Lipid Membranes (2010)
  • Author(s): M.Yamazaki
  • Organizer: ISSP International Workshop of Soft Matter Physics
  • Place of Presentation: IPMU Lecture Hall at Univ.Tokyo, Kashiwa(招待講演)
  • Year and Date: 2010-08-24
• [Presentation] A subcritical half-pore formation in lipid vesicles induced by antimicrobial peptide (2010)
  • Author(s): V.Levadny
  • Organizer: ISSP International Workshop of Soft Matter Physics
  • Place of Presentation: IPMU Lecture Hall at Univ.of Tokyo, Kashiwa
  • Year and Date: 2010-08-23
• [Presentation] Kinetic of low-pH-induced transformation of bilayer membrane to bicontinuous cubic phases in dioleoylphosphatidylserine/monoolein membranes (2010)
  • Author(s): M.M.Alam
  • Organizer: ISSP International Workshop of Soft Matter Physics
  • Place of Presentation: IPMU Lecture Hall at Univ.of Tokyo, Kashiwa
  • Year and Date: 2010-08-23
• [Presentation] Kinetic of low-pH-induced transformation of bilayer membrane to bicontinuous cubic phases in dioleoylphosphatidylserine/monoolein membranes (2010)
  • Author(s): M.M.Alam
  • Organizer: International Symposium on Non-Equilibrium Soft Matter 2010
  • Place of Presentation: Nara Prefectural New Public Hall, Nara
  • Year and Date: 2010-08-18
• [Presentation] Kinetic pathway of antimicrobial peptide magainin 2-induced pore formation in lipid membranes (2010)
  • Author(s): M.Yamazaki
  • Organizer: International Symposium on Non-Equilibrium Soft Matter 2010
  • Place of Presentation: Nara Prefectural New Public Hall, Nara
  • Year and Date: 2010-08-18
• [Presentation] Mechanism of subcritical pore formation in charged lipid vesicles induced by antimicrobial peptide magainin 2 (2010)
  • Author(s): V.Levadny
  • Organizer: International Symposium on Non-Equilibrium Soft Matter 2010
  • Place of Presentation: Nara Prefectural New Public Hall, Nara
  • Year and Date: 2010-08-18
• [Presentation] Interaction of lipopolysaccharide with lipid membranes (2010)
  • Author(s): J.M.Alam
  • Organizer: International Symposium on Non-Equilibrium Soft Matter 2010
  • Place of Presentation: Nara Prefectural New Public Hall, Nara
  • Year and Date: 2010-08-18
• [Presentation] Protein toxin lysenin-induced pore formation in lipid membranes : the single GUV method study (2010)
  • Author(s): R.Takagi
  • Organizer: International Symposium on Non-Equilibrium Soft Matter 2010
  • Place of Presentation: Nara Prefectural New Public Hall, Nara
  • Year and Date: 2010-08-17
• [Presentation] 抗菌ペプチド-マガイニン2の脂質膜中でのポア形成のキネティックスパスウエイ (2010)
  • Author(s): 山崎昌一
  • Organizer: ナノ学会第8回大会
  • Place of Presentation: 岡崎コンファレンスセンター(岡崎)
  • Year and Date: 2010-05-13
• [Presentation] 生体膜のトポロジー変化のダイナミクスとメカニズム (2010)
  • Author(s): 山崎昌一
  • Organizer: 特定研究「ソフトマター物理」AO2班・班会議
  • Place of Presentation: お茶の水女子大学
  • Year and Date: 2010-03-26
• [Presentation] 抗菌ベプチド・マガイニン2が膜中に誘起するボア形成のキネティックスパスウエイ (2010)
  • Author(s): 山崎昌一
  • Organizer: 特定領域「バイオ操作」第8回公開シンポジウム
  • Place of Presentation: 九州大学医学部百年講堂
  • Year and Date: 2010-03-11
• [Presentation] 単一GUV法を用いた生体膜の機能・ダイナミクスの解析(招待講演) (2010)
  • Author(s): 山崎昌一
  • Organizer: 大阪大学 情報科学研究科専攻 ERATO四方研究室セミナー
  • Place of Presentation: 大阪大学
  • Year and Date: 2010-02-22
• [Presentation] 単一GUV法を用いた生体膜の機能・ダイナミクスの解析(招待講演) (2009)
  • Author(s): 山崎昌一
  • Organizer: 名古屋大学 応用物理学教室 談話会
  • Place of Presentation: 名古屋大学
  • Year and Date: 2009-12-17
• [Presentation] 抗菌性ペプチドによる脂質膜中のボア形成の素過程とそのメカニズム:単一GUV法による研究 (2009)
  • Author(s): 山崎昌一
  • Organizer: 特定研究「ソフトマター物理」第3回公開シンポジウム
  • Place of Presentation: 京都大学・船井哲良記念講堂
  • Year and Date: 2009-11-21
• [Presentation] The size of the pore in lipid membranes induced by antimicrobial peptide magainin 2 (2009)
  • Author(s): M.Yamazaki
  • Organizer: 2009 IEEE International Symposium on Micro-NanoMechatronics and Human Science (MHS2009)
  • Place of Presentation: Nagoya
  • Year and Date: 2009-11-10
• [Presentation] Kinetic pathway for the antimicrobial peptide magainin 2-induced pore formation in lipid membranes (2009)
  • Author(s): M.Yamazaki
  • Organizer: 3^<rd> International Symposium on Nanomedicine (ISNM2009-2)
  • Place of Presentation: Okazaki
  • Year and Date: 2009-11-05
• [Presentation] The mechanism of the epigallocatechin gallate-induced burst of giant unilamellar vesicles (2009)
  • Author(s): Y.Tamba
  • Organizer: 日本生物物理学会第47回年次大会
  • Place of Presentation: 徳島 (アスティ徳島)
  • Year and Date: 2009-11-01
• [Presentation] Lysenin-Induced Pore Formation in Lipid Membranes in Liquid-Ordered Phase : the Single GUV Method Study (2009)
  • Author(s): M.Yamazaki
  • Organizer: 日本生物物理学会第47回年次大会
  • Place of Presentation: 徳島 (アスティ徳島)
  • Year and Date: 2009-10-30
• [Presentation] The Size of the Pore in Lipid Membranes Induced by Antimicrobial Peptide Magainin 2 (2009)
  • Author(s): H.Ariyama
  • Organizer: 日本生物物理学会第47回年次大会
  • Place of Presentation: 徳島 (アスティ徳島)
  • Year and Date: 2009-10-30
• [Presentation] Mechanism of Subcritical Pore Formation in Charged Lipid Vesicles Induced by Antimicrobial Peptides (2009)
  • Author(s): V.Levadny
  • Organizer: 日本生物物理学会第47回年次大会
  • Place of Presentation: 徳島 (アスティ徳島)
  • Year and Date: 2009-10-30
• [Presentation] Antimicrobial peptide magainin 2-Induced Pore Formation in Lipid Membranes : the Single GUV Method Study (招待講演) (2009)
  • Author(s): Masahito Yamazaki
  • Organizer: International Symposium on Innovative Nanoscience of Supermolecular Motor Proteins Working in Biomembranes (INSMP)
  • Place of Presentation: Shirankaikan at Kyoto University, Kyoto
  • Year and Date: 2009-09-10
• [Presentation] 抗菌ペプチド:マガイニン2が脂質膜中に形成するポアの大きさ (2009)
  • Author(s): 山崎昌一
  • Organizer: 特定領域「ソフトマター物理」・第4回領域研究会
  • Place of Presentation: 北海道大学・学術交流会館
  • Year and Date: 2009-07-01
• [Remarks]
  • URL: http://www.ipc.shizuoka.ac.jp/~spmyama/index.html
• [Remarks]
  • URL: http://www.ipc.shizuoka.ac.jp/~spmyama/index.html