Fatty acids are very soluble in glycerol or the PEGs.
Most proteins tolerate high concentrations (30-50%) glycerol and PEGs.�
So, it is quite possible to make a solution that will allow the protein
crsytal and the ligand to both be in the same solution.
One of my former colleagues was in ch
Hi Frank,
heme becomes near completely insoluble when it dimerizes. Fatty acids
are also very insoluble. It is quite hard to get them into HSA, as far
as I remember.
Cheers,
Tim
On Wed, 23 Jun 2021 10:33:42 +0100 Frank von
Delft wrote:
> And then of course, you need to decide whether you at al
And then of course, you need to decide whether you at all care to know
anything about a compound that is so insoluble that it needs that kind
of treatment :)
On 23/06/2021 09:52, hoh wrote:
Hi
As total insolubility does not exist, I regularly use another method,
which is to deposit a gr
Dear Gourab,
Sorry for the delayed response, I missed your message earlier.
Can you grow the protein crystals easily (do you have lots of them)?
Is your ligand soluble in PEG, MPD, or Solutol?
Often, you can exchange the crystal solution without destroying the
crystal. But, you have to matc
These are my go-to review articles for ideas for getting a ligand into a
crystal.
https://scripts.iucr.org/cgi-bin/paper?S0907444906047020
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5297911/
I have also seen a talk where someone successfully dissolved their
water-insoluble ligand in Octan-1-
Dear Gourab,
You might just want to add the ligand in pure, solid form to the edge of the
crystallization drop. This worked in our case beautifully (see Angew. Chem.
Int. Ed. 2012, 51, 3354). We had to soak it for 4 weeks.
Good luck!
Best regards
Bernhard
I tried sokaing ,high concentration of DMSO is effecting crystal, so I
tried other contidtion where 10%DMSO is there. Got 1.9 A data with protein
co crystalization with 10mM ligand. But ligand occupancy is not visble.
I got the KD value from ITC, with reverse titration. With 10uM ligand in
cell an
Hello Gourab,
DMSO is not the only possible solvent- there are others to try.
I don't want to sound negative, but 40 micromolar is not a very tight binding
compound. It would also depend on how that binding constant was measured- how
did someone get enough in solution to measure that?
Best of lu
