Hi again,

    I guess this is only a partial summary, since I still don't understand
all the issues this question raises.

Pavel Afonine reported that his extensive tests of the PDB reveals that
reproducing R values from models with TLS ADP's is a wide-spread and
serious problem.  The principal problems (IMHO) are

    1) Incorrect or illegal TLS definitions in the REMARK.

    2) Some files list in the ATOM "B" column the residual B after TLS
       has been accounted for while others list the total B (TLS and
       residual).  There is no clear indication in the PDB file which
       interpretation is being used.

Tassos, Eleanor, and others recommended taking the TLS definition from
the PDB header and running zero cycles of unrestrained refinement in
Refmac to get it to calculate R factors and Maps w/o the need to define
ideal geometry for co-factors.  I have yet to see this work, however
(See below)

Ulrich Baumann wrote to tell me of two of his PDB's that he knows will
give back the reported R values.  They are 2qua and 2qub.

I grabbed 2qua from the RCSB server, extracted the TLS groups with CCP4i,
and found that the TLS definitions were incorrect.  There is one polypeptide
in this model and three TLS groups.  The first and third group did not
have a residue range, while the second group defined a residue range in
the middle of the peptide.  I made the assumption that the first and
third TLS groups were intended to cover the beginning and end of the
peptide and corrected the .tls file.

I loaded this into Refmac and asked for zero cycles of unrestrained
refinement and got an R value of 19.4%.  The PDB file says it should
be 17.3%.  I then asked Refmac to run 10 cycles of TLS and 10 cycles
of restrained refinement and got an R value of 17.5%.  Good enough.

 From this result I infer that Refmac is unable to calculate the original
ADP's given this PDB file and TLS definition.  It can reconstruct them
via refinement, basically ignoring the B values of the PDB file.

This particular PDB entry appears to contain in its "B" column the
residual B's.

I also tried entry 2qub, but with less luck.  This model has seven
peptides and 30 TLS groups.  The first seven TLS groups defined in
the header of the PDB cover each of the seven chains, while the other
23 groups had no residue range.  I can guess that the intension was
to have five TLS groups for each of the seven chains, but without
additional information from Dr. Baumann, I'm unable to even start
trying to reproduce R values and calculate maps.

So...  1) Pavel is correct, there are many clear errors in the TLS
REMARKs of PDB entries.  2) It seems necessary to ask Refmac to
recreate the ADP description for a PDB entry from scratch, assuming
the TLS group definition can be deduced from the PDB header.  This,
currently, requires refinement which requires .cif's for the unusual
groups.

If CCP4I could ask Refmac to perform only TLS/B refinement, holding
positions fixed, the need for detailed .cifs would be greatly reduced.
I have no desire to move the atoms anyway.

Better yet, if someone could find out what Refmac is expecting to find
in its starting PDB (what it wants in the "B" column) one could add
a tool to CCP4I that could convert a PDB entry to what Refmac wants
w/o refinement.  Since there appear to be two varieties of entries
one could try both possibilities and choose the one with the lowest
R value.

I have to close with additional problems, I'm afraid.  I can't run
the required refinement on 1nkz to test TLS/B refinement but
I have tried it on 3bsd, where I have a good .cif for the Bchl-a
groups.  When I pull out the TLS definition, and perform 10 cycles
of TLS and 10 cycles of restrained refinement I get an R value of
20.2% while the entry asserts that the correct value is 17.8%.  The
final TLS parameters look, by eye, pretty similar to the deposited
ones, so I don't know what is going on here.

Dale Tronrud



Dale Tronrud wrote:
> Hi,
> 
>    I am looking over a number of models from the PDB but have been
> unable to reproduce the R-factors for any model that was refined
> with Refmac and contains TLS parameters.  I usually can't get within
> 5% of the reported value.  On the other hand, I usually do pretty
> well for models w/o TLS.
> 
>    An example is the model 1nkz.  The PDB header gives an R value
> of 17% but even when I use tlsanal in CCP4i to generate a PDB with
> anisotropic B's that mimic the TLS parameters I get an R value of
> 22.4% using SFCheck.  (I'm not implying that I suspect any problem
> with 1nkz, in fact I have every reason to believe this is the great
> model its published stats indicate.)
> 
>    I've found a CCP4 BB letter that stated that SFCheck does not
> pay attention to anisotropic B's but that letter was dated 2002.
> I hope this limitation has been removed, or at least the output
> would mention this limitation.
> 
>    Setting up a refinement in Refmac involves a large overhead,
> since even for zero cycles of refinement the program insists on
> a complete stereochemical definition for the strange and wondrous
> groups in this model.  I would just like to verify the R factor
> and calculate a proper map for inspection in Coot.  Since I have
> many models I would like to look at, I would like a simple procedure.
> 
>    I did set up a Refmac run for another model, for which I do
> have all the .cif's required, but even after refinement I was not
> close to the reported R.
> 
>    I see that the models I'm interested in are not present in the
> Electron Density Server, so I suspect I'm not alone in fighting
> this battle.
> 
> Any advice would be appreciated,
> Dale Tronrud