We only notice the dust when we let pucks thaw while still assembled. We had
always assumed that the dust was due to oxidation (different types of metals in
contact with the water from condensation). We have not seen the dust since we
began taking apart pucks and removing pins prior to thawing t
Hi,
a likely scenario is that your mutant crystallises in the same
conformation/packing as the wild-type protein, and this conformation is good
for ligand binding. In solution, your mutant protein may be more flexible/open
than the wild-type and affinity hence lower. We see this quite often. Var
As other said, having co-crystal doesn’t mean you will have measurable
affinity. Further no heat in ITC experiments doesn’t mean no binding. Which
buffer you used during previous ITC experiment? If heat in the previous
experiment (wild type) was due to heat of ionization and you removed the
res
As other said, having co-crystal doesn’t mean you will have measurable
affinity. Further no heat in ITC experiments doesn’t mean no binding. Which
buffer you used during previous ITC experiment? If heat in the previous
experiment (wild type) was due to heat of ionization and you removed the
res
Dear Monika,
If your protein-ligand interaction is at millimolar, you need to increase
injection volume in order to see heat change. Also, you can do the experiment
at low temperature such as 10 degree Celsius. You can follow this paper where I
have crystallized weak binding ligand and compleme
Alexis
This is a very useful summary.
You say you were not convinced by Marin's derivation in 2005. Are you convinced
now and, if not, why?
My interest in this is that the FSC with half bit thresholds have the danger of
being adopted elsewhere because they are becoming standard for protein
str
Hi Monika,
If the protein cocrystalize with ligand doesn't mean it interacts with
protein.
You have provided insufficient information here. Does the ligand is bound
to mutant protein with atleast 2 or 3 hydrogen bonds (how many residues are
mutated?) ? If no that means, it is just cocrystalizing b
Beside no change in enthalpy, was there no change in the entropy and
affinity?
Pankaj Kumar Chauhan, PhD
Assistant Professor
Ramalingaswami Fellow
Department of Biochemistry
Jamia Hamdard University (Institute of Eminence by Govt. of India)
Hamdard Nagar, Delhi, India 110062
Skype: pankajimtech
P
Fully agree with what others have said. When ’the dust’ happened to us we had
to replace that particular shipping dewar as T was not holding for very long.
Best wishes
Roberto
On 22 Feb 2020, at 00:33, Emilia C. Arturo (Emily)
mailto:ecgart...@gmail.com>> wrote:
Hi All,
We have been noticing
