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Originally published as Biophys J. BioFAST on May 20, 2005.
doi:10.1529/biophysj.105.059550
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Biophysical Journal 89:1046-1054 (2005)
© 2005 The Biophysical Society

Photocurrents Generated by Bacteriorhodopsin Adsorbed on Nano-Black Lipid Membranes

Christian Horn and Claudia Steinem

Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, Regensburg, Germany

Correspondence: Address reprint requests to Claudia Steinem, Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, 93040 Regensburg, Germany. Tel.: 49-941-943-4548; Fax: 49-941-943-4491; E-mail: claudia.steinem{at}chemie.uni-regensburg.de.

Purple membranes were adsorbed on freestanding lipid bilayers, termed nano-black lipid membranes (nano-BLMs), suspending the pores of porous alumina substrates with average pore diameters of 280 nm. Nano-BLMs were obtained by first coating the upper surface of the highly ordered porous alumina substrates with a thin gold layer followed by chemisorption of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol and subsequent addition of a droplet of 1,2-diphytanoyl-sn-glycero-3-phosphocholine and octadecylamine dissolved in n-decane onto the hydrophobic submonolayer. By means of impedance spectroscopy, the quality of the nano-BLMs was verified. The electrical parameters confirm the formation of single lipid bilayers with high membrane resistances covering the porous matrix. Adsorption of purple membranes on the nano-BLMs was followed by recording the photocurrents generated by bacteriorhodopsin upon continuous light illumination. The membrane system exhibits a very high long-term stability with the advantage that not only transient but also stationary currents are recordable. By adding the proton ionophore carbonyl cyanide-m-chlorophenylhydrazone the conductivity of the nano-BLMs increases, resulting in a higher stationary current, which proves that proton conductance occurs across the nano-BLMs.




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E. K. Schmitt, M. Vrouenraets, and C. Steinem
Channel Activity of OmpF Monitored in Nano-BLMs
Biophys. J., September 15, 2006; 91(6): 2163 - 2171.
[Abstract] [Full Text] [PDF]


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