Dear Robert,

In addition to the remarks and suggestions by others: How do your electron 
density maps look like? Do they look remotely reasonable, or do they look like 
they had gone through a meat grinder? If they look remotely ok, you could also 
try manual rebuilding, pruning etc. However, if they look chopped, there is no 
sense in trying to (re)build anything in it, either manually or automatically. 
I guess you tried all permutations and variations of P2221, with zero, one and 
two screw axes in all positions?

Best,
Herman

Von: CCP4 bulletin board <[email protected]> Im Auftrag von Robert S 
Phillips
Gesendet: Donnerstag, 18. Juni 2020 15:01
An: [email protected]
Betreff: [EXTERNAL] [ccp4bb] Molecular replacement problem


EXTERNAL : Real sender is 
[email protected]<mailto:[email protected]>

I've been pulling out my hair with this for a few months now.  I have data sets 
to 2.6 A for a new enzyme in the aminotransferase superfamily.  Unfortunately, 
the closest structure is only 25% identity.  MR with PHASER using the monomer 
was a complete failure.  Since the minimum structure of enzymes in the family 
is a dimer (the active site is formed at the monomer-monomer interface), I used 
dimers for MR with PHASER.  Most of the results were marginal, but one looks 
good.  However, it will not refine.  Everything I have done with this solution 
has failed, simulated annealing, morphing, etc., it will not refine; AUTOBUILD 
and BUCCANEER with it do not give any useable models, since they have poor 
statistics and low completeness.  The output from PHASER is below.

** SINGLE solution

** Solution written to PDB file:  DGL_phaser.1.pdb
** Solution written to MTZ file:  DGL_phaser.1.mtz
   Solution annotation (history):
   SOLU SET  RFZ=18.9 TFZ=23.0 PAK=0 LLG=193 TFZ==8.3 LLG=994 TFZ==20.1 PAK=0 
LLG=994 TFZ==20.1
   SOLU SPAC P 2 2 21
   SOLU 6DIM ENSE ense_1 EULER  360.0    0.0    0.0 FRAC -0.00 -0.00 -0.00 BFAC 
-0.12 MULT 2 #TFZ==20.1
   SOLU ENSEMBLE ense_1 VRMS DELTA -3.5403 #RMSD  2.08 #VRMS  0.89

With LLG = 994 and TFZ = 20.1, isn't this a real solution?

Robert S. Phillips
Professor of Chemistry and of Biochemistry and Molecular Biology
University of Georgia
Athens, GA 30602
Phone: (706) 542-1996
Fax: (706) 542-9454
E-mail: [email protected]<mailto:[email protected]>
Web:  
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