[phenixbb] Failed to refine B factors of some residues and ligands

Pavel Afonine pafonine at lbl.gov
Tue Nov 15 17:50:14 PST 2016


Hi Christian,

somehow this escaped my attention... so was this solved in the end? If 
not please send me files and I will find a solution.

Pavel

On 10/31/16 10:55, Christian Roth wrote:
>
> Not more than 5 I'm afraid. I don't know if it would find the right 
> B's using 10 cycles and more, but that also increases the running time 
> quite considerably even on some of our faster machines.
>
> Best wishes
>
> Christian
>
>
> Am 31.10.2016 um 16:46 schrieb Dorothee Liebschner:
>> Hi Christian,
>>
>> OK, it looks you found a way to prevent the positive density by using 
>> "realistic" starting B values (of the order of that of surrounding 
>> protein atoms).
>> Did you also try to increase the number of macro-cycles?
>>
>> Best wishes,
>>
>> Dorothee
>>
>> On Mon, Oct 31, 2016 at 9:11 AM, Christian Roth 
>> <christianroth034 at gmail.com <mailto:christianroth034 at gmail.com>> wrote:
>>
>>     Hi Dorothee
>>
>>     to answer your questions
>>
>>     The resolution is about 2.0 Ang and the average B around 55. The
>>     Bfactor from the ligand was set to the default of 20 by Coot when
>>     placing the ligands.
>>
>>     It seems that setting the B-value to the average B of the
>>     structure nearly solves the problem (most of the ligand fine,
>>     just parts with unusuallly high values). Setting it to a bit
>>     higher values 70 for a start and the ligand refines fine.
>>
>>     I noticed that in the first macrocycle with and initial factor of
>>     20, the B's are nearly unchanged (20 +- 2) Red diff density
>>     appears as expected. In the second cycle The minimizer let the
>>     B-factors "explode" to about 200 and that basically is the
>>     overkill and it never recovers.
>>
>>     HTH
>>
>>     Cheers
>>
>>     Christian
>>
>>     Am 31.10.2016 um 15:59 schrieb Dorothee Liebschner:
>>>     Hi Christian,
>>>
>>>     Could you answer the following questions:
>>>
>>>     - what is the resolution of the data set?
>>>     - what is average B of the protein?
>>>     - what are the starting B values for the ligand?
>>>     - did you try lowering starting B for the ligand and repeat the
>>>     refinement? Is this behavior reproducible with different
>>>     refinement options?
>>>
>>>     Best wishes,
>>>
>>>     Dorothee
>>>
>>>     On Sun, Oct 30, 2016 at 11:06 AM, Christian Roth
>>>     <christianroth034 at gmail.com <mailto:christianroth034 at gmail.com>>
>>>     wrote:
>>>
>>>         Hi Phenix Team,
>>>
>>>         regarding that ligand issue in this thread, we observed the
>>>         same with a ligand in one of our structures now. After
>>>         refinement the ligand is surrounded by pos diff density and
>>>         the B-Factors are around 200 (way above any useful range in
>>>         that case).
>>>
>>>         The cif was generated using elbow and phenix runs with
>>>         options Real space refinement, TLS on, individual B, and
>>>         optimizing weights. Nothing crazy.
>>>
>>>         Could you find a solution for the previous case and give us
>>>         some tips?
>>>
>>>
>>>         Cheers
>>>
>>>
>>>         Christian
>>>
>>>
>>>
>>>
>>>         Am 06.09.2016 um 01:44 schrieb Pavel Afonine:
>>>>         Hi Tongqin,
>>>>
>>>>         I second Dorothee's point that we need more information to
>>>>         resolve this problem. If you still have this problem please
>>>>         send me inputs files (data, model, .eff from last
>>>>         refinement, as well as ligand .CIF files if any) and I will
>>>>         look into this once I have files.
>>>>
>>>>         Pavel
>>>>
>>>>         On 8/31/16 09:29, Dorothee Liebschner wrote:
>>>>>         Hi Tongqin,
>>>>>
>>>>>         It is a bit difficult to diagnose with the information you
>>>>>         provided. Could you please answer the following questions?
>>>>>
>>>>>         - Did you also try to reset the B-factors to similar
>>>>>         values than neighboring atoms?
>>>>>         F.ex. the average B-factor in the model could be 50 A**2,
>>>>>         but in the ligand region, it could be lower, let's say 20
>>>>>         A**2. Then the starting value using average B is quite far
>>>>>         from the likely B-factor of the ligand.
>>>>>
>>>>>         - What refinement strategy do you apply? How many
>>>>>         macro-cycles? Do you use any non-default parameters for
>>>>>         B-factor refinement?
>>>>>
>>>>>         - Which Phenix version are you using?
>>>>>
>>>>>         Best wishes,
>>>>>
>>>>>         Dorothee
>>>>>
>>>>>         On Wed, Aug 31, 2016 at 7:12 AM, Zhou, Tongqing (NIH/VRC)
>>>>>         [E] <tzhou at mail.nih.gov <mailto:tzhou at mail.nih.gov>> wrote:
>>>>>
>>>>>             Dear All,
>>>>>
>>>>>             I am refining a structure with diffraction to 2.2A
>>>>>             resolution and 95 % overall completeness. Now the Rs
>>>>>             are at 18% and 23%, respectively. But the program
>>>>>             failed to refine B factors for several ligands, atoms
>>>>>             had high B factors while showing positive Fo-Fc map
>>>>>             around them. Occupancy was set to 1 and I also tried
>>>>>             to reset B to mean B of the whole PDB. Any of you
>>>>>             seeing this phenomenon? Thanks!
>>>>>
>>>>>             Shown below is HEPES that has high B factor and
>>>>>             positive density around it after refinement:
>>>>>
>>>>
>>>>
>>>>
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