Item
2: Topic discussion: NCS
SUMMARY: NCS is part of single crystal
X-ray diffraction data and a useful addition to the
crystallographic symmetry. All experimental data should be used
for derivation of experimental results. NCS contains "error" that
should be treated properly with the use of a weighting scheme. CNS
and SHELXL use different approaches in structure refinement with
NCS restraints.
Experts' views from outside the group:
http://www.ysbl.york.ac.uk/ccp4bb/2002/msg00083.html
http://www.ysbl.york.ac.uk/ccp4bb/2002/msg00091.html
That's
quite interesting. I have not used TNT with NCS constraints or restraints. With
X-plor/CNS you have both options as well. In the case of strict NCS, only one
molecule is refined and then rotation/ translation matrix(es) will be applied
to generate one or more copies. This does not allow any deviation between the
copies. If deviations are anticipated, one should use NCS restraints and set
appropriate weights. In this case, equivalent sets of atoms are superimposed
and their coordinates are restrained to be close to the average position E(ncs)
= SUM w(x-x(mean))**2 ( with x being the vector x,y,z). However, this cannot
account for domain movements around hinges. If the molecule contains domains,
NCS-related molecules might in fact display different rotations around those
hinges. One can then define those regions separately. Furthermore, the backbone
atoms may obey the NCS very well, whereas the side chains, especially those on
the surface, usually don't. Figuring out which atoms belong to a NCS group can
be a tedious job and one is sooner or later ready to compromise (i.e. give up
NCS restraints partially ). In contrast, Shelxl has a better way to deal with
NCS by setting up 'local NCS' restraints. The NCSY instruction selects NCS
related atoms but sets up 1-4 distance restrains internally. This automatically
allows for domain rotations without any problem. In my experience, it works
very well and is quite flexible in that it allows you to select main chain or
side chain atoms or both depending on the degree of similarity. By the way, it
seems that X-plor can in fact apply local NCS similar to Shelx (unless I am
mistaken), but it is much less obvious. One would have to use NOE energy terms
and use distance restraints. For a crystallographer, this is a bit unfamiliar. (Lothar Esser, NCI)
The most powerful fashion
to use noncrystallographic symmetry is to reduce the number of parameters in
your model with constraints. The correspondence of the separate images of the
molecule is assumed to be perfect and the model does not allow any variation
between copies. Basically the number of parameters in the model decreases by
the number of images in the asymmetric unit. When there are real differences
between the copies of the molecule, you do not want to use the
noncrystallographic symmetry as constraints. To allow the data to tell you
where the differences are you must allow some variation in your model. These
requirements require that you incorporate the non-crystallographic symmetry as
restraints. You will have to choose a weight to set the relative importance of
these observations to your other observations. (TNT Users' Guide)
Yes, I use NCS whenever it
is observable. Why? NCS is in fact experimental data at either atomic (for
identical copies of the molecule) or lower (for non-identical copies)
resolutions, which contributes in both structure solution and refinement. For
structure refinement, NCS data at atomic resolution can be used as constraints
to powerfully reduce the number of refined parameters, whereas lower-
resolution NCS data can be used as restraints for all or selected region(s) of
the molecule with a proper weighting scheme. One way of choosing the region(s)
and weights is to start the refinement with NCS restraints for the entire
molecule and tighter restraints and then follow the indications of Fo-Fc.
(Xinhua Ji, NCI)
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