While the B value in the starting model was fine.  I did drop it to a 
reasonable value and it now seems to be behaving, was thinking about trying 
that before I posted.  Since this was an  MR determination I did check and the 
starting model B value was actually smaller then the surrounding residues.  I 
guess some thing went odd with the first round of refinement.  

Thank you for the suggestions.

Len


> On Aug 10, 2022, at 10:25 AM, Dale Tronrud <[email protected]> wrote:
> 
> 
>   A large B value with positive difference density sure implies a convergence 
> problem with the refinement.  Was the B value extreme in your starting model? 
>  (A starting B that is wildly too large or too small at the start may cause 
> it to become trapped in the refinement.) Maybe if you rerun your refinement 
> with a moderate starting value for the B you will end up a more sensible 
> result.
> 
>   The only other way to end up with a parameter that directly conflicts with 
> the difference density is a bad restraint, but that doesn't sound likely 
> based on your description.
> 
> Dale Tronrud
> 
> On 8/10/2022 7:59 AM, Thomas, Leonard M. wrote:
>> Hello All,
>> I have run into something odd.  In working on a structure for one of the 
>> groups I work with regularly, on one of the cystine residues I have a very 
>> large positive density peak at the sulfur position. The B value is 
>> approximately 4 times the other values in the residue and on other cystine 
>> residues.  The overall structure has 2 molecules in the asymmetric unit  and 
>> the corresponding cystine  on the other monomer is behaving as I would 
>> expect.   There are no disulfides in the structure.
>> The data were collected on 9-2 at SSRL and all three of the data sets we 
>> collected show the same thing, all data go to about 2.2 angstroms.  We are 
>> trying to determine the ligand binding in the molecule but this cystine is 
>> not involved in ligand binding.  In house and other synchrotron data from 
>> previous protein preps and data collection runs of the same molecule grown 
>> in very similar condition and crystallized in the same space group have the 
>> residue behaving normally.
>> I am open to any ideas as to what may be going on as I am rather puzzled by 
>> this.
>> Thanks for any input,
>> Len Thomas
>> Leonard Thomas, Ph.D.
>> Biomolecular Structure Core, Director
>> Oklahoma COBRE in Structural Biology
>> Price Family Foundation Institute of Structural Biology
>> University of Oklahoma
>> Department of Chemistry and Biochemistry
>> 101 Stephenson Parkway
>> Norman, OK 73019-5251
>> Office: (405)325-1126
>> [email protected] <mailto:[email protected]>
>> http://www.ou.edu/structuralbiology/cobre-core-facilities/mcl 
>> <http://www.ou.edu/structuralbiology/cobre-core-facilities/mcl>
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