Elucidating mechanisms


09-May-2015 20:38

Upon adsorbing to the electrode surface (see ), it is crucial that the protein molecules retain their native structure and properties.This is easily confirmed for a redox enzyme because its catalytic properties should be retained.The direct electrochemistry of redox proteins and enzymes, protein film voltammetry, is becoming established as a method which is able to provide precise and comprehensive information on complicated reaction mechanisms [1], [2] and [3].In protein film voltammetry (PFV), the protein is adsorbed directly onto the electrode, removing the necessity for it to diffuse to the surface.

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Protein film voltammetry, the direct electrochemistry of redox enzymes and proteins, provides precise and comprehensive information on complicated reaction mechanisms.By controlling the driving force for a reaction (using the applied potential) and monitoring the reaction in real time (using the current), it allows thermodynamic and kinetic information to be determined simultaneously.It is less easily confirmed for a non-catalytic redox protein, but one would hope that at least one property of the protein, such as the reduction potential or spectroscopic signature of a cofactor, may measured both in solution and on the electrode surface and compared .Although large shifts in potential should not be tolerated without reason, reduction potentials often do shift slightly upon adsorption (by 20 m V or so) reflecting the altered environment or the preferential adsorption of one oxidation state, and perturbations may be observed when the protein is adsorbed within a film of co-adsorbate because it is embedded within the electrical double layer .

Two challenges are inherent to protein film voltammetry: (i) to adsorb the protein or enzyme in a native and active configuration on the electrode surface, and (ii) to understand and interpret voltammetric results on both a qualitative and quantitative level, allowing mechanistic models to be proposed and rigorous experiments to test these models to be devised.This review focuses on the second of these two challenges.



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