Asked • 03/15/19

Is the EC50 of an activating protein for an enzyme a good indicator for the binding affinity Kd?

We work with a membrane protein system where measuring the affinity between the enzyme and the upstream activating protein has been difficult, and when measured in detergent solution, it is almost 100 fold lesser (ie ~100nM) whereas the EC50 in an enzymatic assay using vesicles in ~1-2nM. Would it be reasonable to say that the "real" affinity is ~1nM than 100s of nM based on the biochemical assay?Alternately, is there a documented system where a huge discrepancy exists between measurements from direct binding and biochemical assays?

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