I have used potassium ferricyanide (which is likely the most common) extensively in the past, but I am curious to know if there are other oxidants that work well. I need to generate the oxidized cluster in my protein (which purifies in the [4Fe4S]2+ redox state), and certain downstream applications preclude the use of any iron-based oxidants.
I would love to know suggestions if you have any!
For those wondering, my protein is unstable (in vitro) when its cluster is in the oxidized state (I actually use a proteolytically-stable domain of a larger enzyme as a model system that becomes unstable when oxidized). The time it takes to remove the ferricyanide usually leads to precipitation of all of the oxidized protein (which I've verified by EPR). Similar patterns are observed with the complete enzyme in absence of its substrate. While it would be simple to be able to just collect data for this protein in the presence of its substrate, It's crucial that I also collect data without the enzyme's desired substrate.
Also check https://onlinelibrary.wiley.com/doi/full/10.1111/j.1365-2958.2006.05028.x
The [4Fe-4S]2+ clusters of dehydratases are rapidly damaged by univalent oxidants, including hydrogen peroxide, superoxide, and peroxynitrite. The loss of an electron destabilizes the cluster, causing it to release its catalytic iron atom and converting the cluster initially to an inactive [3Fe-4S]1+ form. https://pubmed.ncbi.nlm.nih.gov/15308657/
Kindly check https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/iron-sulfur-cluster
Also check https://onlinelibrary.wiley.com/doi/full/10.1111/j.1365-2958.2006.05028.x
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