I worked with several viral cystene proteases. I do not recommend using
basic pH for them dring purification or storage or crystallization. Keep it
below 7, because the active site cysteine oxydazes very easily. The higher
the pH, the easier the oxydation. PH 10 can also cause hydrolysis of some
I fitted manually the chondroitin sulfate into my complex (3c9e.pdb and 3h7d)
using program coot. The chondroitin-sulfate model was used from the PDB. I used
CNS for the refinement of the first structure, and phenix for the second
structure. Phenix was more convenient with this ligand as it was
>> b. the kD in solution is indeed higher that the concentration you
>> used for MALLS
>> c. The constructs are not well chosen for some reason
>>
>> You could use AUC to detect kD as high as ~100uM, depending on the
>> concentration of the start sample of co
The multimeric state depends on a protein concentration. You can get any
multimer to dissociate if you dilute it to low enough concentration. If
your complex is a homodimer, then Kdiss=[complex]/[monomer]^2. Let's say
your Kdiss~10^(-3)M, and your protein concentration is ~10^(-4)M, then
[com
Why do you need a fusion protein? Can you use His-tag directly? If you have to
have a fusion protein, use one that has no hydrophobic patches on the surface,
because otherwise they will be glued to each other. For separation of your
mixture that you have now try dilute it as much as possible ( b
There are Rpim and Rrim, Rpim is sqrt(1/(N-1)) and is usually small and Rrim
(or Rmeas)=sqrt(N/(N-1)) and is large. I usually go with I/sigma cutoff.
Maia
- Original Message -
From: "Edward A. Berry"
To:
Sent: Thursday, April 22, 2010 11:59 AM
Subject: Re: [ccp4bb] Rsym problems..
Dear all,
I have just subscribed to the CCP4BB. The description for this BB is "The
CCP4BB mailing list is for discussions on the use of the CCP4 suite, and
macromolecular crystallography in general. "
So, please, give us all possible solutions to crystallographic problems,
including protein e
