Jacob,
In case if the hint that I sent yesterday was not clear, below is the solution 
for the equation
Kd=[P][L]/[PL] 

in terms of ligand occupancy:

O=[ PL]/[Po]= 1/(Kd/L+1)

You see, it does not depend on [Po]

Alex

On Jun 26, 2011, at 10:05 AM, aaleshin wrote:

> The concentration of a protein in a crystal [Po] and the volume of a crystal 
> V are needed only to calculate the total amount of a ligand [Lo] required for 
> soaking.
> [Lo]  > [Po]*V
> 
> The occupancy of the active sites in a crystal will depend only on the ligand 
> concentration in solution and Kd. It does not depend on protein concentration 
> in the crystal.
> 
> Indeed:
> Kd=[P][L]/[PL]
> 
> Assuming total concentration of the protein = Po, Kd= 1mM and S= 1 mM, the 
> active site occupancy will be:
> 
> 1= P/Po-P;
> 
> P/Po=1/2
> 
> So the concentration of the ligand in solution should be >>Kd to get the full 
> occupancy.
> 
> Alex
> 

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