It is possible that your PO4 has its atoms labeled with the wrong
chirality.  Yes, I know that PO4 is not chiral when you ignore hydrogen
atoms and single/double bonds but adding labels creates an unnatural
chriality.  Try your refinement again after switching the labels on two
oxygen atoms.

Dale Tronrud

On 6/22/2015 7:48 AM, ansuman biswas wrote:
> Dear CCP4 users,
> 
> I am working on a protein from a hyperthermophilic archaeon.
> 
> I have collected mutliple X-Ray datasets, both from home source and
> synchrotron and always found a clear density for tetrahedral geometry,
> co-ordinated by two histidines and one lysine. 
> 
> I tried fitting phosphate there, but its geometry always gets distorted
> after each refinement cycle (Refmac 5.8.0073). Also I found some short
> contacts between the coordinated residues and phosphate which were very
> difficult to remove.
> 
> I am attaching a figure with the density and phosphate. 
> 
> Kindly suggest -
> 1. if this may be a possible modification of any of the associated
> residues, and the code of the modified residue to be used.
> 
> 2. If the ligand requires separate restraints during refinement, I am
> using the "restrained refinement" option available at the top of the GUI
> for refmac.
> 
> Thanking you,
> yours sincerely,
> Ansuman Biswas,
> PhD student,
> Dept. of Physics,
> IISc
> 

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