Hi Kaibo,

It is normal to find more sites than are in the sequence, both due to multiple conformations of SeMet side chains and due to identifying other anomalous scatterers as Se.  It is fine for you to edit these sites for refinement based on chemical plausibility.   You can also use the difference in f" values for different scatterers to give clues as to the identity of any particular scatterer.  This can be done in phenix.refine.

All the best,
Tom T


On Mon, Jul 9, 2018 at 3:32 PM, Pavel Afonine <pafonine@lbl.gov> wrote:
Hi Kaibo,

2. The 2Fo-Fc map and AutoBuild solution looked fine, but the Fo-Fc map is kinda noisy with positive peaks showing up within the main-chain density which causes troubles for refinement.

what contouring level you use to see Fo-Fc map? Note, usual "3 sigma" is not universally optimal but depends on solvent content. Do you still see lots of artifacts at say 0.35 or 0.4e/A**3 (mean solvent density) ?

Pavel

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Hello,

I tried to use AutoSol to solve a SAD date set. The space group is P212121, with two proteins per ASU each containing one SeMet. However, the Hyss would always find 4~5 extra sites per ASU besides the two SeMet residues.

So my questions are:
a. Is this normal to have so many more sites identified? Or what might be the problem?
b. Could I remove those extra Se sites during refinement if the occupancy is low or doesn't make sense biologically?

Here are some details:
1. Most of the extra sites are around 20% occupancy and not within the range as alternate SeMet side-chain positions. One of them even overlapped with the position where should be a ligand phosphate.

2. The 2Fo-Fc map and AutoBuild solution looked fine, but the Fo-Fc map is kinda noisy with positive peaks showing up within the main-chain density which causes troubles for refinement.

3. No twining was detected for the dataset. Several space groups have been tried (including no screw axis) and all of the solutions contained more sites than there should be.
 
Please let me know if you encountered similar problems before. Thanks a lot!

Best regards
Kaibo
 

Kaibo Zhang
PULSe, Purdue University
Hockmeyer Hall, Room 321
240 S. Martin Jischke Drive, West Lafayette, IN 47907
Telephone: 765-337-1955
E-mail: zhang644@purdue.edu




--
Thomas C Terwilliger
Laboratory Fellow, Los Alamos National Laboratory
Senior Scientist, New Mexico Consortium
100 Entrada Dr, Los Alamos, NM 87544
Tel: 505-431-0033