Hmmm - TLS may not be applied to the waters . The interaction between 10
fold ncs snd TLS snd water positions could have strange outcomes! Like Kyle
I wonder e what happens if you deactivate both those options?

On Tue, 6 Jan 2026 at 18:14, Kay Diederichs <
[email protected]> wrote:

> Dear Kyle,
>
> interesting finding - worth looking into!
>
> According to Wlodawer et al (2024) (
> https://journals.iucr.org/m/issues/2024/06/00/be5300/index.html) ,"An
> analysis of the contents of the PDB indicated that the expected ratio of
> the number of water molecules to the number of amino-acid residues exceeds
> 1.5 in atomic resolution structures, decreasing to 0.25 at around 2.5 Å
> resolution."
> So I'd expect your structure to have around 800 residues, is that about
> right?
>
> A lot may depend on the details of data processing ... which program; is
> there anisotropy? How did you determine/perform the high-resolution cutoff?
> Could you go to higher resolution? Do you have other datasets from the same
> project - do they show the same behaviour?
>
> I can imagine refinement/water-adding workflows where some atoms have
> surprising B-values e.g. TLS refinement mistakenly leading to anisotropic
> B-values, or waters being deleted and re-added with a default B-value of
> 16. Could you give details?
> As a simple test, just remove any lines from your PDB file except the
> ATOM, HETATM, DISU and the CRYST1 and END records. Then refine with a
> standard Refmac protocol and check the B-values afterwards.
>
> HTH,
> Kay
>
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