A metal-supported biomimetic micromembrane allowing the recording of single-channel activity and of impedance spectra of membrane proteins

被引:15
作者
Becucci, Lucia [1 ]
D'Amico, Massimo [2 ]
Daniele, Salvatore [3 ]
Olivotto, Massimo [2 ]
Pozzi, Andrea [1 ]
Guidelli, Rolando [1 ]
机构
[1] Univ Florence, Dept Chem, I-50019 Florence, Italy
[2] Univ Florence, Dept Expt Pathol & Oncol, I-50134 Florence, Italy
[3] Univ Venice, Dept Phys Chem, I-30123 Venice, Italy
关键词
Tethered bilayer lipid membrane; OmpF porin; Single-channel current; Electrochemical impedance spectroscopy; BILAYER-LIPID MEMBRANES; MICROELECTRODE ARRAYS; ION-TRANSPORT; VALINOMYCIN; THIOLIPIDS; ELECTRODES; MONOLAYERS; MOLECULES; MELITTIN; KINETICS;
D O I
10.1016/j.bioelechem.2009.08.007
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A novel tethered bilayer lipid micromembrane (tBL mu M) was prepared and characterized. It consists of a mercury cap electrodeposited on a platinum microelectrode, about 20 pm in diameter. The micromembrane was prepared by tethering to the mercury cap a thiolipid monolayer and by then self-assembling a lipid monolayer on top of it. The thiolipid consisted of a disulfidated tetraoxyethylene hydrophilic spacer covalently linked to two phytanyl chains. Upon incorporating OmpF porin in the tBL mu M, its single-channel activity was recorded by the patch-clamp technique, and its particular features described. An electrochemical impedance spectrum of the tBL mu M incorporating OmpF porin is also reported. To the best of our knowledge, this tBL mu M is the first metal-supported biomimetic micromembrane capable of incorporating non-engineered channel proteins in a functionally active state from their detergent solutions, and of allowing the recording of single-channel activity and of impedance spectra of these proteins via ion translocation into the hydrophilic spacer. The limited spaciousness of the spacer prevents a statistical analysis based on current-amplitude or blockage-time histograms. Nonetheless, the robustness, stability, ease of preparation and disposability of the present tBL mu M may open the way to the realization of a channel-protein microarray platform allowing a high throughput drug screening. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:176 / 180
页数:5
相关论文
共 32 条
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