金(111)电极表面浮动磷脂双层膜的原位偏振红外反射光谱研究(英文)
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摘要
应用电化学原位偏振红外反射光谱法研究了构建于金(111)电极表面的浮动磷脂双层膜.金电极表面先自组装一层巯基葡萄糖单层来增加表面的亲水性,浮动磷脂双层膜通过LB-LS技术构建在巯基葡萄糖单层上.双层膜由双肉豆蔻磷脂酰胆碱(DMPC)、胆固醇和神经节苷脂GM1构成.GM1分子中的糖链可以物理吸附在巯基葡萄糖表面,在双层膜和基底间形成一个富含水的隔层.红外光谱表明浮动双层膜中的DMPC分子比传统的支撑双层膜中的DMPC分子有更强的水合作用,证实了双层膜和基底间水层的存在.该浮动双层膜更接近于实际的生物膜体系,并且在金电极表面有宽的电位区间,非常适于进一步的离子通道蛋白质研究.
In situ polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) was used to study the structure of a DMPC+cholesterol+GM1 floating bilayer lipid membrane (fBLM) at a Au(111) surface.1-thio-β-Dglucose (β-Tg) was self-assembled onto the Au electrode to increase the overall hydrophilicity of the surface.The fBLM was then deposited on the β-Tg self-assembled monolayer (SAM) using a combination of Langmuir-Blodgett/Langmuir-Schaefer (LB/LS) techniques.The carbohydrate headgroups of the GM1 molecules were physically adsorbed to the β-Tg SAM forming a water rich cushion between the fBLM and the modified gold substrate.The PM-IRRAS spectra indicate that the DMPC molecules within the fBLM are more hydrated than previous studies involving supported bilayer lipid membranes (sBLM) where the membrane is directly adsorbed onto the surface.The tilt angle of the DMPC acyl chains in the fBLM is smaller than that of the sBLM composed of similar components.The results from this work confirmed that the fBLM is stable over a wide range of electrode potentials and that a water rich region is present between the bilayer and gold electrode surface.The addition of this water region more closely mimics the natural environment of a biological membrane making the fBLM a desirable candidate for future in situ studies involving transmembrane proteins.
In situ polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) was used to study the structure of a DMPC+cholesterol+GM1 floating bilayer lipid membrane (fBLM) at a Au(111) surface.1-thio-β-Dglucose (β-Tg) was self-assembled onto the Au electrode to increase the overall hydrophilicity of the surface.The fBLM was then deposited on the β-Tg self-assembled monolayer (SAM) using a combination of Langmuir-Blodgett/Langmuir-Schaefer (LB/LS) techniques.The carbohydrate headgroups of the GM1 molecules were physically adsorbed to the β-Tg SAM forming a water rich cushion between the fBLM and the modified gold substrate.The PM-IRRAS spectra indicate that the DMPC molecules within the fBLM are more hydrated than previous studies involving supported bilayer lipid membranes (sBLM) where the membrane is directly adsorbed onto the surface.The tilt angle of the DMPC acyl chains in the fBLM is smaller than that of the sBLM composed of similar components.The results from this work confirmed that the fBLM is stable over a wide range of electrode potentials and that a water rich region is present between the bilayer and gold electrode surface.The addition of this water region more closely mimics the natural environment of a biological membrane making the fBLM a desirable candidate for future in situ studies involving transmembrane proteins.
引文
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[39]Li M,Chen M,Sheepwash E,et al.AFM studies of sol-id-supported lipid bilayers formed at a Au(111)elec-trode surface using vesicle fusion and a combination of Langmuir-Blodgett and Langmuir-Schaefer techniques[J].Langmuir,2008,24(18):10313-10323.
[40]Kycia A H,Sek S,Su Z F,et al.Electrochemical and STM Studies of1-thio-β-d-glucose self-assembled on a Au(111)electrode surface[J].Langmuir,2011,27(21):13383-13389.
[41]Raguse B,Braach-Maksvytis V,Cornell B A,et al.Tethered lipid bilayer membranes:Formation and ionic reservoir characterization[J].Langmuir,1998,14(3):648-659.
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[45]Lee D C,Durrani,A A,Chapman D.A difference in-frared spectroscopic study of gramicidin A,alamethicin and bacteriorhodopsin in perdeuterated dimyris-toylphosphatidylcholine[J].Biochimica et Biophysica Acta,1984,769(1):49-65.
[46]Brosseau C L,Bin X,Roscoe S G,et al.Electrochemi-cal and PM-IRRAS characterization of DMPC+choles-terol bilayers prepared using Langmuir-Blodgett/Lang-muir-Schaefer deposition[J]Journal of Electroanalytical Chemistry,2008,621(2):222-228.
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[50]Bin X,Zawisza I,Lipkowski J.Electrochemical and PM-IRRAS studies of the effect of the static electric field on the structure of the DMPC bilayer supported at a Au(111)electrode surface[J].Langmuir,2005,21(1):330-347.
[51]Zawisza I,Bin X,Lipkowski J.Potential-driven struc-tural changes in Langmuir-Blodgett DMPC bilayers de-termined by in situ spectroelectrochemical PM IRRAS[J].Langmuir,2007,23(9):5180-5194.
[52]Umemura J,Kamata T,Takenaka T.Quantitative evalu-ation of molecular orientation in thin Langmuir-Blod-gett films by FT-IR transmission and reflection-absorp-tion spectroscopy[J].The Journal of Physical Chemistry,1990,94(1):62-67.
[53]Hauser H,Pascher I,Pearson R H,et al.Preferred conformation and molecular packing of phos-phatidylethanolamine and phosphatidylcholine[J].Biochimica et Biophysica Acta,1981,650(1):21-51.
[54]Aussenac F,Laguerre M,Schmitter J M,et al.Detailed structure and dynamics of bicelle phospholipids using selectively deuterated and perdeuterated labels.2H NMR and molecular mechanics study[J].Langmuir,2003,19(25):10468-10479.
[55]Lewis R N A H,McElhaney R N.Components of the carbonyl stretching band in the infrared spectra of hy-drated1,2-diacylglycerolipid bilayers:A reevaluation[J].Biophysical Journal,1994,67(6):2367-2375.
[56]Fringeli U P.The structure of lipids and proteins studied by attenuated total reflection(ATR)infrared spec-troscopy.II.Oriented layers of a homologous series:Phosphatidylethanolamine to phosphatidylcholine[J].Z Naturforsch C,1977,32(1/2):20-45.
[57]Fringeli U P,Gunthard H H.Molecular biology,bio-chemistry and biophysics[M].Grell E,Ed.,Springer Verlag:Berlin,1981:270-332.
[58]Fringeli U P.A new crystalline phase of L-alpha-di-palmitoyl phosphatidylcholine monohydrate[J].Bio-physical Journal,1981,34(2):173-1887.
[59]Hubner W,Blume A.Interactions at the lipid-water in-terface[J].Chemistry and Physics of Lipids,1998,96(1/2):99-123.
[2]Alberts B,Johnson A,Lewis J,et al.Molecular biology of the cell[M].5th Edition.New York:Garland Science,2007:617-650.
[3]Papahadjopoulos D,Watkins J C.Phospholipid model membranes.I.Structural characteristics of hydrated liq-uid crystals[J].Biochimica et Biophysica Acta,1967,135(4):624-638.
[4]Hauser H,Phillips M C,Stubbs M.Ion permeability of phospholipid bilayers[J].Nature,1972,239:342-344.
[5]Nagle J F,Tristram-Nagle S.Structure of lipid bilayers[J].Biochimica et Biophysica Acta,2000,1469(3):159-195.
[6]Sackmann E.Supported membranes:Scientific and prac-tical applications[J].Science,1996,271(5245):43-48.
[7]Castellana E T,Cremer P S.Solid supported lipid bilay-ers:From biophysical studies to sensor design[J].Surface Science Reports,2006,61(10):429-444.
[8]Mendelsohn R,Mao G,Flach C R.Infrared reflection-ab-sorption spectroscopy:Principles and applications to lipid-protein interaction in Langmuir films[J].Biochimica et Biophysica Acta,2010,1798(4):788-800.
[9]Lee C S,Bain C D.Raman spectra of planar supported lipid bilayers[J].Biochimica et Biophysica Acta,2005,1711(1):59-71.
[10]Lirtsman V,Ziblat R,Golosovsky M,et al.Surface-plasmon resonance with infrared excitation:Studies of phospholipid membrane growth[J].Journal of Applied Physics,2005,98(9):093506.
[11]Dabkowska A P,Fragneto G,Hughes A V,et al.Specu-lar Neutron reflectivity studies of the interaction of cy-tochrome c with supported phosphatidylcholine bilayers doped with phosphotidylserine[J].Langmuir,2009,25(7):4203-4210.
[12]Nomura K,Inaba T,Morigaki K,et al.Interaction of lipopolysaccharide and phospholipid in mixed mem-branes:Solid-state 31 P-NMR spectroscopic and micro-scopic investigations[J].Biophysical Journal,2008,95(3):1226-1238.
[13]Miller C E,Majewski J,Watkins E B,et al.Probing the local order of single phospholipid membranes using grazing incidence X-ray diffraction[J].Physical Review Letters,2008,100(5):058103.
[14]Tamm L K,McConnell H M.Supported phospholipid bilayers[J].Biophysical Journal,1985,47(1):105-113.
[15]Tamm L K.Lateral diffusion and fluorescence micro-scope studies on a monoclonal antibody specifically bound to supported phospholipid bilayers[J].Biochem-istry,1988,27(5):1450-1457.
[16]Groves J T,Ulman N,Cremer P S,et al.Substrate-membrane interactions:Mechanisms for imposing pat-terns on a fluid bilayer membrane[J].Langmuir,1998,14(12):3347-3350.
[17]Kuhner M,Tampe R,Sackmann E.Lipid mono-and bi-layer supported on polymer films:Composite poly-mer-lipid films on solid substrates[J].Biophysical Jour-nal,1994,67(1):217-226.
[18]Guidelli R,Aloisi G,Becucci L,et al.Bioelectrochem-istry at metal/water interfaces[J].Journal of Electroana-lytical Chemistry,2001,504(1):1-28.
[19]Lang H,Duschl C,Vogel H.A new class of thiolipids for the attachment of lipid bilayers on gold surfaces[J].Langmuir,1994,10(1):197-210.
[20]Rossi C,Chopineau J.Biomimetic tethered lipid mem-branes designed for membrane-protein interaction stud-ies[J].European Biophysics Journal,2007,36(8):955-965.
[21]Naumann R,Jonczyk A,Kopp R.et al.Incorporation of membrane proteins in solid-supported lipid layers[J].Angewandte Chemie International Edition,1995,34(18):2056-2058.
[22]Kycia A H,Wang J P,Merrill A R,et al.Atomic force microscopy studies of a floating-bilayer lipid membrane on a Au(111)surface modified with a hydrophilic monolayer[J].Langmuir,2011,27(17):10867-10877.
[23]Fa guy P W,Richmond W N.Real-time polarization modulation infrared spectroscopy applied to the study of water and hydroxide ions at electrode surfaces[J].Jour-nal of Electroanalytical Chemistry,1996,410(1):109-113.
[24]Barth A.Infrared spectroscopy of proteins[J].Biochimi-ca et Biophysica Acta,2007,1767(9):1073-1101.
[25]Vie V,Legardinier S,Chieze L,et al.Specific anchoring modes of two distinct dystrophin rod sub-domains inter-acting in phospholipid Langmuir films studied by atom-ic force microscopy and PM-IRRAS[J].Biochimica et Biophysica Acta,2010,1798(8):1503-1511.
[26]Brosseau C L,Leitch J,Bin X,et al.Electrochemical and PM-IRRAS a glycolipid-containing biomimetic membrane prepared using Langmuir-Blodgett/Lang-muir-Schaefer deposition[J].Langmuir,2008,24(22):13058-13067.
[27]Chen M,Li M,Brosseau C L,et al.AFM Studies of the effect of temperature and electric field on the structure of a DMPC-cholesterol bilayer supported on a Au(111)electrode surface[J].Langmuir,2009,25(2):1028-1037.
[28]Ohta Y,Yokoyama S,Sakai H,et al.Membrane proper-ties of mixed ganglioside GM1/phosphatidylcholine monolayers[J].Colloids and Surfaces B:Biointerfaces,2004,33(3/4):191-197.
[29]Li N,Zamlynny V,Lipkowski J,et al.In situ IR re-flectance absorption spectroscopy studies of pyridine adsorption at the Au(110)electrode surface[J].Journal of Electroanalytical Chemistry,2002,43(524-525):43-53.
[30]Green M J,Barner B J,Corn R M.Real-time sampling electronics for double modulation experiments with Fourier transform infrared spectrometers[J].Review of Scientific Instruments,1991,62(6):1426-1430.
[31]Buffeteau T,Desbat B,Turlet J M.Polarization modula-tion FT-IR spectroscopy of surfaces and ultra-thin films:Experimental procedure and quantitative analysis[J].Applied Spectroscopy,1991,45(3):380-389.
[32]Buffeteau T,Desbat B,Blaudez D,et al.Calibration procedure to derive IRRAS spectra from PM-IRRAS spectra[J].Applied Spectroscopy,2000,54(11):1646-1650.
[33]Zamlynny V,Lipkowski J.Diffraction and spectroscop-ic methods in electrochemistry[M]//Alkire R,Kolb D M,Lipkowski J,et al.Ed.Weinheim:Wiley-VCH,2006,9:315-376.
[34]Zamlynny V.PhD thesis[D].University of Guelph,2002.
[35]Palik E.Ed.Handbook of optical constants of solids II[M].Academic Press:San Diego,1998.
[36]Bertie J E,Ahmed M K,Eysel H H.Infrared intensities of liquids.5.Optical and dielectric constants,integrated intensities,and dipole moment derivatives of H 2 O and D 2 O at22℃[J].The Journal of Physical Chemistry,1989,93(6):2210-2218.
[37]Lipert R J,Lamp B D,Porter M D.Modern techniques in applied molecular spectroscopy[M]//Mirabella F M.Ed.New York:John Wiley&Sons,Inc.,1998:83-126.
[38]Leonard A,Escrive C,Laguerre M,et al.Location of cholesterol in DMPC membranes.A comparative study by neutron diffraction and molecular mechanics simula-tion[J].Langmuir,2001,17(6):2019-2030.
[39]Li M,Chen M,Sheepwash E,et al.AFM studies of sol-id-supported lipid bilayers formed at a Au(111)elec-trode surface using vesicle fusion and a combination of Langmuir-Blodgett and Langmuir-Schaefer techniques[J].Langmuir,2008,24(18):10313-10323.
[40]Kycia A H,Sek S,Su Z F,et al.Electrochemical and STM Studies of1-thio-β-d-glucose self-assembled on a Au(111)electrode surface[J].Langmuir,2011,27(21):13383-13389.
[41]Raguse B,Braach-Maksvytis V,Cornell B A,et al.Tethered lipid bilayer membranes:Formation and ionic reservoir characterization[J].Langmuir,1998,14(3):648-659.
[42]Burgess I,Szymanski G,L,M,et al.Electric field-driv-en transformations of a supported model biological membrane—an electrochemical and neutron reflectivity study[J].Biophysical Journal C,2004,86(3):1763-1776.
[43]Gennis R B.Biomembranes,molecular structure and function[M].New York:Springer-Verlag,1989.
[44]Liang C Y,Lytton M R.Infrared spectra of crystalline and stereoregular polymers[J].Journal of Polymer Sci-ence,1962,61(172):S45-S48.
[45]Lee D C,Durrani,A A,Chapman D.A difference in-frared spectroscopic study of gramicidin A,alamethicin and bacteriorhodopsin in perdeuterated dimyris-toylphosphatidylcholine[J].Biochimica et Biophysica Acta,1984,769(1):49-65.
[46]Brosseau C L,Bin X,Roscoe S G,et al.Electrochemi-cal and PM-IRRAS characterization of DMPC+choles-terol bilayers prepared using Langmuir-Blodgett/Lang-muir-Schaefer deposition[J]Journal of Electroanalytical Chemistry,2008,621(2):222-228.
[47]Nabet A,Auger M,Pezolet M.Investigation of the tem-perature behavior of the bands due to the methylene stretching vibrations of phospholipid acyl chains by two-dimensional infrared correlation spectroscopy[J].Applied Spectroscopy,2000,54(7):948-955.
[48]Allara D L,Swalen J D.An infrared reflection spec-troscopy study of oriented cadmium arachidate mono-layer films on evaporated silver[J].The Journal of Phys-ical Chemistry,1982,86(14):2700-2704.
[49]Allara D L,Nuzzo R G.Spontaneously organized molecular assemblies.1.Formation,dynamics,and physical properties of n-alkanoic acids adsorbed from solution on an oxidized aluminum surface[J].Langmuir,1985,1(1):45-52.
[50]Bin X,Zawisza I,Lipkowski J.Electrochemical and PM-IRRAS studies of the effect of the static electric field on the structure of the DMPC bilayer supported at a Au(111)electrode surface[J].Langmuir,2005,21(1):330-347.
[51]Zawisza I,Bin X,Lipkowski J.Potential-driven struc-tural changes in Langmuir-Blodgett DMPC bilayers de-termined by in situ spectroelectrochemical PM IRRAS[J].Langmuir,2007,23(9):5180-5194.
[52]Umemura J,Kamata T,Takenaka T.Quantitative evalu-ation of molecular orientation in thin Langmuir-Blod-gett films by FT-IR transmission and reflection-absorp-tion spectroscopy[J].The Journal of Physical Chemistry,1990,94(1):62-67.
[53]Hauser H,Pascher I,Pearson R H,et al.Preferred conformation and molecular packing of phos-phatidylethanolamine and phosphatidylcholine[J].Biochimica et Biophysica Acta,1981,650(1):21-51.
[54]Aussenac F,Laguerre M,Schmitter J M,et al.Detailed structure and dynamics of bicelle phospholipids using selectively deuterated and perdeuterated labels.2H NMR and molecular mechanics study[J].Langmuir,2003,19(25):10468-10479.
[55]Lewis R N A H,McElhaney R N.Components of the carbonyl stretching band in the infrared spectra of hy-drated1,2-diacylglycerolipid bilayers:A reevaluation[J].Biophysical Journal,1994,67(6):2367-2375.
[56]Fringeli U P.The structure of lipids and proteins studied by attenuated total reflection(ATR)infrared spec-troscopy.II.Oriented layers of a homologous series:Phosphatidylethanolamine to phosphatidylcholine[J].Z Naturforsch C,1977,32(1/2):20-45.
[57]Fringeli U P,Gunthard H H.Molecular biology,bio-chemistry and biophysics[M].Grell E,Ed.,Springer Verlag:Berlin,1981:270-332.
[58]Fringeli U P.A new crystalline phase of L-alpha-di-palmitoyl phosphatidylcholine monohydrate[J].Bio-physical Journal,1981,34(2):173-1887.
[59]Hubner W,Blume A.Interactions at the lipid-water in-terface[J].Chemistry and Physics of Lipids,1998,96(1/2):99-123.