Dear Ruby,
Since you give very little details, I can only give a few random tips.
- Does your protein aggregate? Did you check light scattering? If it does
aggregate, you may have to try adding things like detergents or glycerol.
- Try orthologues, add ligands (was mentioned by others as well).
- Heat the protein for a short time to say 70°C to get rid of misfolded and
contaminating proteins.
- Is your protein part of a multiprotein complex? In that case it might have
hydrophobic surfaces which were not optimized be nature for solubility, but as
protein-protein interfaces. In this case you may try cocrystallization with one
or more partner(s). You may also try to predict hydrophobic surface residues
and make some mutations to enhance solubility.
- Try a completely different protein or try to get ligand complexes from a
crystallized protein, so you will at least have something to write in your
thesis. Otherwise you risk that your time will be up and you have nothing to
write about.
Good luck!
Herman
________________________________
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of
ruby singh
Sent: Friday, May 04, 2012 9:10 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb]
Dear all,
Im a PhD student who started working on Protein crystallization. Its
been years im trying to get any crystal of that protein. I have tried all
crystallization screens available.
but no results. My protein is highy thermostable(till 80C) and pH
stable(1 to 13)...plz if anyone is having any idea or trick let me knw.
