so if I have for example an inhibitor that shows only 20% of inhibition (that being maximum) of some enzyme
Can I still do the kinetics study?
It may perhaps be useless to do it - but still, could it be done?
Are there some limitations to maximum inhibition percentage to even start with kinetic study...
Hi there,
Do you mean that the maximal effect of the inhibitor at the maximal dose toward a given dose of protein is the inhibition of 20% of the enzyme activity? If yes your molecule is rather a negative allosteric modulator. The binding of the modulator on its binding site provokes a rearrangement of the active site which becomes catalytically less active (but still active). And of course you may calculate the Kd for the enzyme /modulator interaction (it will be the concentration required for half of the maximum effect, in your case the dose required to observe 10% inhibition).
I agree with Dominique, but I will add that if the maximal inhibition of 20% is really due to insolubility of the compound, then you may reach the wrong conclusion about what it does.
Ok. Dominique it is as you say. But Adam is also right. I didn't mention solubility. Higher concentration is not soluble.
I have sesquiterpene hydrocarbon that i am trying to test against acetyl/butyrylcolinesterase. I have been using Ellman assay and EtOH to dissolve it. Tried also Acetone and DMSO - but no good. Any advices...
In general what would be considered as a good solvent for this kind of a system?
For such an insoluble substance, I think you would have to use a micellar detergent to keep it in solution. The problem with this approach is that it is difficult to define the functional concentration.
I am now reading my previous answer. I have been using EtOH (and it did disolve in it) and I got some good effects on AChE but not on BuChE at low concentrations (1,47mM-final) because higher ones are (blurry/cloudy). Could it be concluded that it was in contact with the enzyme enough to exhibit the effect or ...?
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