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Hi all,
This is a question to which everyone would like to know the answer (i.e.,
there is no correct answer), but I'd like to know if anyone has a strong
opinion on this:
I have been trying to crystallize a fairly insoluble protein. After some
engineering and buffer optimization I've achieved a 5-fold increase in
solubility. However, still no luck on getting crystals. I am thinking about
cloning from a different organism, and have looked at all homologs in the
Pfam database (I know thermophiles are popular for crystallization trials).
I have ruled out a number of isoforms based on their stability index,
expressability (?!), and availability; and have been looking into using a
program which estimates the solubulity of the peptide based on it's sequence
(any specific recommendations?).
Does anyone have any further suggestions towards being smart about which
organism to clone from?
Thanks!
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