Carles, a few points to consider:
- you might be comparing B-factors coming from local atomic vibrations alone (residual B, or whatever it's called), and the total atomic B-factor, that includes all components: Btotal = Btls + Bresidual +... . (for comprehensive review on this topic see: TLS from fundamentals to practice, Crystallography Reviews, (2013), 19:4, 230-270) - there may be a number of valid reasons for the mean B-factor to deviate from Wilson B. - as Ethan mentioned, obviously your goal is not to preserve the original B-factors. All the best, Pavel On Thu, Jun 5, 2014 at 8:51 AM, Carles Palanca i García < [email protected]> wrote: > Hello, > > > I'm almost done refining a protein-DNA complex at 3A, but when I apply > TLS (Refmac+TLSanl to include the ANISOU component) the B factors increase > from 50-60 to 100-110 A2. The crystal is a bit special since it has 80% of > solvent and the Wilson B-factor is 87 A2. When I compare my Bfactors with > other structures of similar resolution using Phenix Validation, my data is > clearly an outlier. > > > To perform the TLS refinement I have generated .tlsin files for 2,4 and > 6 groups with the TLSmd server and used them in refmac jobs consisting of > 20 cycles of TLS refinement+50 cycles of restrained refinement. This way > the R-factor and R-free values have drop 1-2% (before it was 22.3/24.1). > The three different options suffer from the same problem regarding high > B-factors after TLSanl. Am I doing something wrong during the TLS > refinement? What can I do to preserve the original Bfactors? > > > Thanks in advance, > > Carles Palanca. > >
