Hi,
a few days ago, in searching results from the Brenda web site, i found that compound "A" can inhibit competitively enzyme "B". The compound "A" has similar structure with substrate "C" and high similarity score at 0.83 in KEGG SIMCOMP (ranked at top 8). A hydroxy- on end of carbon chain is just difference between the A and C.
I'm looking for some article paper or method to remove or avoid this competitive inhibition, Can u recommend me?
Thank you.
I assume you mean to remove the inhibitor from your enzyme preparation.
If it is a competitive inhibitor and if your substrate or product is not too expensive, add a large mount of either and dialyze your enzyme solution and, if you can, also to the dialysis buffer..
The substrate or product will compete out the inhibitor from the binding site and be removed during the dialysis. After this first dialysis, you would have to dialyze the enzyme preparation at least twice against buffer without substrate or product, to get rid of these, as well.
Hi there,
It may depend on the inhibitor affinity. According to what you say, A and C affinities shouldn't be too different therefore chasing A with C is a good option. Direct dialysis of your enzyme prep contaning the inhibitor versus buffer is the easiest and cheapest option (provided BA complex dissociation rate is high enough).
If the inhibitor is not bound too tightly (Kd in µM), another simple way to remove it is by passing the protein through a gel filtration desalting column.
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