Hi Satya Dev,

You can feed the mtz output of SCALA to Phenix Xtriage and then see the
presence of t-NCS and/or any other crystal pathologies. Another thing you
can do is to merge and scale the data in P222 and then let the Phaser
decide the best space group. Again, I am curious, when you ran pointless,
did you check the absence conditions and the probability of assigning all
the three screw axes?

Good luck!
Sudipta.

On Sat, Mar 18, 2017 at 5:42 AM, Eleanor Dodson <[email protected]>
wrote:

> You dont say whether there is Non cryst translation - that will be
> reported at various stages - the pointless/aimless/ctruncate task gives it.
>
> But if it exists and the translation ihas a component of .5 along any
> axis, that makes the SG estimate a bit uncertain - the absences could be
> due to the NX translation.
>
> And even if the SG is correct - which likely after solving the MR with the
> newest PHASER which tests carefully = then you will have zones with low
> intensities, and those reflections always have a higher r factor of course.
>
> You could let Arp/Warp or Buccaneer rebuild starting from your existing
> model? That is a verification that your solution is essentially right
>
> Eleanor
>
>
>
> On 18 March 2017 at 10:28, Isupov, Michail <[email protected]> wrote:
>
>> Hi,
>>
>> I have seen cases where in a correct space group
>> 'R-work and R-free values 0.25 and 0.32 respectively'
>> at 2 A resolution sound like not too bad values.
>> In some of such cases when data from a different crystal
>> in the same space group was available R-factors were much lower
>> when the structure was refined against the new crystal data.
>> I guess this phenomenon could be due to uneven freezing of the first
>> crystal,
>> or inconsistent degree of disorder between crystals.
>>
>> In other projects high R-factor values (e.g. FreeR around 33% at 2.1 A
>> resolution)
>>  are consistent through a range of crystals
>> even when the refinement is in P1, although the map quality is good enough
>> to see cofactors and to build  the missing parts of the structure (30% of
>> residues).
>> The disorder seems to be an intrinsic property of such crystal form.
>>
>> I do not know how to approach publishing these results since most
>> referees will argue
>> that such R-factors may be acceptable at 4A resolution but not close to 2
>> Angstrom.
>>
>> Best wishes,
>>
>> Misha Isupov
>> ________________________________________
>> From: CCP4 bulletin board [[email protected]] on behalf of Randy
>> Read [[email protected]]
>> Sent: Saturday, March 18, 2017 9:29 AM
>> To: [email protected]
>> Subject: Re: [ccp4bb] No improvement in R-factor after Refmac.
>>
>> Hi,
>>
>> I was just going to make the same point!  The only thing to add is that,
>> if there really is translational NCS (which is certainly possible with 4
>> copies in the a.u.), then it’s essential both to account for it (which
>> current versions of Phaser should do automatically, if you search for all 4
>> copies in one job) and to try all possible space groups.  The situation
>> Craig describes, in which it’s not immediately obvious whether your crystal
>> has a crystallographic 2(1) and a pseudosymmetric non-crystallographic
>> 2-fold or the reverse, is not uncommon.  However, we’ve found that the
>> likelihood score accounting for the effect of tNCS is pretty good at
>> discriminating the two possibilities.
>>
>> Best wishes,
>>
>> Randy Read
>>
>> -----
>> Randy J. Read
>> Department of Haematology, University of Cambridge
>> Cambridge Institute for Medical Research    Tel: +44 1223 336500
>> Wellcome Trust/MRC Building                         Fax: +44 1223 336827
>> Hills Road
>> E-mail: [email protected]
>> Cambridge CB2 0XY, U.K.
>> www-structmed.cimr.cam.ac.uk
>>
>> > On 18 Mar 2017, at 06:12, CRAIG A BINGMAN <[email protected]> wrote:
>> >
>> > You really need to approach such situations with caution.  Examination
>> of the relatively small number of axial reflections probably show that
>> there might be twofold screw axes in all three directions.  But a
>> non-crystallographic microscopic translation of nearly 0.5 in the direction
>> of a crystallographic axis will give the same pattern of strong and weak
>> reflections as a crystallographic twofold screw axis.  If I were you, I
>> would be very sure to try molecular replacement in all possible
>> orthorhombic space groups.  Several programs, including Phaser, will
>> organize that exhaustive search across all eight possibilities for you.
>> >
>> >> On Mar 17, 2017, at 11:56 PM, Polisetty Satya Dev <[email protected]>
>> wrote:
>> >>
>> >> Hi,
>> >>
>> >> We checked all possible space groups of orthorhombic crystal system
>> using Scala and Pointless but the statistics show that P212121 is the
>> possible space group.
>> >>
>> >> Thank You,
>> >> Satya Dev
>> >>
>> >> On Fri, Mar 17, 2017 at 8:03 PM, Teplyakov, Alexey [JRDUS] <
>> [email protected]> wrote:
>> >> Check the space group. It may be orthorhombic with a pure rotational
>> axis (e.g. P21212) or even monoclinic.
>> >>
>> >>
>> >>
>> >> From: CCP4 bulletin board [mailto:[email protected]] On Behalf Of
>> Polisetty Satya Dev
>> >> Sent: Friday, March 17, 2017 9:51 AM
>> >> To: [email protected]
>> >> Subject: [EXTERNAL] [ccp4bb] No improvement in R-factor after Refmac.
>> >>
>> >>
>> >>
>> >> Dear all,
>> >>
>> >> I solved a structure at 2.0 A resolution with R-work and R-free values
>> 0.25 and 0.32 respectively and I am stuck at Refmac step where there is no
>> further reduction in R-factor.
>> >>
>> >> The above stated values were obtained after several rounds of manual
>> refinement followed by refmac. There are also areas where electron density
>> is missing around peptide backbone in one of the monomer in ASU.
>> >>
>> >> Can anyone please tell me how can I improve the electron density and
>> R-factor.
>> >>
>> >>
>> >> The structure solution was obtained using Phaser MR and here are the
>> data statistics:
>> >>
>> >>
>> >>
>> >> Average unit cell: 81.95, 100.40, 156.96,   90.00, 90.00, 90.00,
>> >> Space group: P212121,
>> >> Completeness 99.5,
>> >> Multiplicity 6.4,
>> >> Four monomers per ASU.
>> >> Solvent content: 47%.
>> >>
>> >> Thank you everyone,
>> >> Satya Dev,
>> >> JNCASR.
>> >>
>> >
>>
>
>

Reply via email to