Protein electrochemistry offers a direct method to identify and characterize biological electron transfer processes, potentially leading to commercial applications such as biosensors and diagnostic tools. However, establishing a biocompatible electrode interface that maintains the native state of the redox protein involves several challenges. In general, membrane proteins require the presence of a phospholipid bilayer to maintain their biological activity. Synthetic `biomimetic’ membranes are widely used to characterize membrane proteins, however they have seldom been applied to measurements of protein redox activity in electrochemical cells due to their inherent insulating property. In this study we demonstrate the use of the phospholipids: PC, PC/PG and PC/PG/cholesterol membrane mixtures on chemically modified (supported) gold electrode surfaces for direct protein electrochemistry. We compare the electrochemical activity of a relatively small, redox active “test protein”, cytochrome c, in the presence and absence of phospholipid on a gold electrode modified with thiol self assembled monolayers, to explore the effect of chain length and composition of the thiol on the charge coupling. Three thiols were investigated as self assembled monolayers on a gold electrode: octanethiol, mercaptopropionic and mercaptoundecanoic acid. We demonstrate here that the charge transfer efficiency of cytochrome c is better in the presence of the membrane and in addition, a superior redox response is obtained with surfaces modified with a thiol functionalised with a carboxylic acid.
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Abbreviations
- MPA:
-
mercaptopropionic acid
- MUA:
-
mercaptoundecanoic acid
- SAM:
-
self-assembled monolayer
- PBS:
-
phosphate buffered saline
- DMPC:
-
1,2-dimyristoyl-l-α-phosphatidylcholine
- DMPG:
-
1,2-dimyristoyl-l-α-phosphatidyl-dl-glycerol
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Acknowledgments
This work was supported by a grant from the National Health and Medical Research Council. The Australian Research Council Discovery and LIEF Grants, Monash University and Victorian Institute for Chemical Sciences are also acknowledged for financial support for purchase of a Q-sense QCM and electrochemical equipment. We thank Dr John Lee for technical assistance with the Scheme.
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On leave from: Research Group on Laser Physics of the Hungarian Academy of Sciences, University of Szeged, Szeged, Hungary.
Australian Peptide Conference Issue.
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Mechler, A., Nawaratna, G., Aguilar, MI. et al. A Study of Protein Electrochemistry on a Supported Membrane Electrode. Int J Pept Res Ther 12, 217–224 (2006). https://doi.org/10.1007/s10989-006-9029-0
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DOI: https://doi.org/10.1007/s10989-006-9029-0