I agree with Herman about shorter soak. In addition, if your crystals can
tolerate any change in pH, you can slow your reaction down by tweaking this
parameter. Another thing you should think about is the kinetics of the
reaction, especially the rate limiting step. If the rate limiting step is
Dear Petr,
Another possibility is that your very good substrate got turned over by the
enzyme and that the 5 atoms with good electron density you see is all that is
left. When you know the enzymatic reaction, this is easy to check. If this is
the case, you should try a short soak (5-30
