Fabrication of Semi-Transparent Gold Electrodes and Modification of the Gold Surface with Self-Assembled Monolayer (SAM)
6:07
Electrochemical Testing of Proteoliposomes Activity
7:46
Detection of Enzymatic Proton Pumping by Fluorescence Microscopy
9:44
Results: Liposome Identification and pH Change
11:35
Conclusion
Transkript
This method allows the study of proton transport activity of respiratory membrane enzymes like cytochrome BO3 in a natural lipid bi-layer environment. The main advantages of this single enzyme technique is the possibility to start and stop enzymat
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Here, we present a protocol to study the molecular mechanism of proton translocation across lipid membranes of single liposomes, using cytochrome bo3 as an example. Combining electrochemistry and fluorescence microscopy, pH changes in the lumen of single vesicles, containing single or multiple enzyme, can be detected and analyzed individually.