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Development of Membrane-Based Biosensors: Measurement of Current from Photocycling Bacteriorhodopsin on Patch Clamp Electrodes

  • Paul Yager
Part of the Advances in Experimental Medicine and Biology book series (AEMB)

Abstract

Our initial work toward developing membrane protein-based biosensors has involved use of bacteriorhodopsin (BR) as a model membrane protein. BR was incorporated into liposomes of a polymerizable lecithin, and was shown to pump protons in response to illumination both before and after polymerization of the lipids1,2. In the work described in this paper, BR was first reincorporated in liposomes of asolectin by cosonication with purple membrane. The liposomes, which sustained the function of the protein, were used to form a monolayer at the air-water interface. This monolayer was transferred as a bilayer onto patch electrode. When illuminated with a pulse of 514.5-nm light the lipid/protein patch produced a current spike into the pipette corresponding to events no later than the generation of the 412-nm intermediate, probably caused by pumping of protons across the patch membrane. The experiment demonstrates not only the extreme sensitivity of amperometric detection, but also a small tendency for membrane proteins to preferentially orient in this configuration.

Keywords

Light Pulse Current Signal Purple Membrane Patch Electrode Current Spike 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Paul Yager
    • 1
  1. 1.Code 6190 Bio/Molecular Engineering BranchNaval Research LaboratoryUSA

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