共 32 条
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.
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页码:176 / 180
页数:5
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