I would also be suspicious of the space group/unit cell if you have 10 mols in 
the A.U. especially if the R-free is unusually high but still reasonable.

Best,
Omid

From: CCP4 bulletin board <[email protected]> On Behalf Of Rafael Marques
Sent: January 7, 2026 8:05 AM
To: [email protected]
Subject: Re: [ccp4bb] Unusually low B-factors for 'waters'

[△EXTERNAL]

Very interesting topic,

Sorry for highjacking a bit the discussion, but I always pay attention to the 
model b-factor instead the individual ones. In this specific case, what 
b-factors should be expected for the water molecules?

Best wishes


______________________________________________________

Rafael Marques da Silva

PhD Student – Structural Biology

University of Leicester

Mestre em Física Biomolecular
Universidade de São Paulo

Bacharel em Ciências Biológicas
Universidade Federal de São Carlos

phone: +44 07861 273773

           "A sorte acompanha uma mente bem treinada"
________________________________________________
________________________________
De: CCP4 bulletin board <[email protected]<mailto:[email protected]>> 
em nome de Clemens Vonrhein 
<[email protected]<mailto:[email protected]>>
Enviado: quarta-feira, 7 de janeiro de 2026 11:21
Para: [email protected]<mailto:[email protected]> 
<[email protected]<mailto:[email protected]>>
Assunto: Re: [ccp4bb] Unusually low B-factors for 'waters'

As an addition to that good idea: you could run BUSTER (via
"aB_autorefine" command-line) starting from an empty model to get
automatic water addition, TLS, occ-refinement, hydrogen handling etc
as a 2nd/3rd opinion. If you need help getting started with BUSTER,
contact us off-list ;-)

Cheers

Clemens

On Tue, Jan 06, 2026 at 12:47:45PM -0800, Pavel Afonine wrote:
> get a second opinion. Phenix, Refmac, or Shelxl are your options. If you
> refined your model with one of them, try another. If you choose
> phenix.refine, let it find/build/refine waters automatically. This way, you
> can rule out a bug in the software you used to obtain this result or
> modeling error. Of course, if this happens with Phenix, feel free to send
> me the files off-list, and I will have a close look!
>
> Good luck!
> Pavel
>
>
> On Tue, Jan 6, 2026 at 6:48 AM Kyle Gregory <
> [email protected]<mailto:[email protected]>>
>  wrote:
>
> > Dear all,
> >
> > I have a 2.6 angstrom structure, which has 10 molecules in the ASU. I have
> > modelled approx 200 water molecules. They fit spherical density nicely and
> > make suitable contacts with the protein. The only problem is nearly every
> > single water molecule has a lower B-factor than the protein atoms they
> > coordinate. E.g protein atoms could be 30/40 while water is 16 angstrom^2.
> >
> > Does anyone have a suggestion as to why, and whether there are some things
> > I should be trouble shooting?
> >
> > Kind regards,
> > Kyle
> >
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--

*--------------------------------------------------------------
* Clemens Vonrhein, Ph.D.     vonrhein AT GlobalPhasing DOT com
* Global Phasing Ltd., 9 Journey Campus, Castle Park
* Cambridge CB3 0AX, UK                   
www.globalphasing.com<http://www.globalphasing.com>
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