It seems to be taken for granted that radiolysis of water generates hydrogen 
gas. I was in a lecture 25 years ago when the speaker said that the gas bubbles 
which appear when a protein crystal that has been in the beam is warmed to room 
temperature is also hydrogen. Seems eminently sensible. However a year or two 
later someone told me that some effort was going into finding out what gas it 
was experimentally. Only gossip and hearsay from long ago and I never heard any 
more about it, sorry ...

Best wishes, Jon Cooper (Emeritus at UCL) [email protected]
Erratum and other hopefully useful things: https://crxp.org.uk

Sent from [Proton Mail](https://proton.me/mail/home) for Android.

-------- Original Message --------
On Monday, 07/20/26 at 18:43 Pavel Afonine 
<[email protected]> wrote:

> Hi Rafael,
>
> Indeed, hydrogen abstraction is an interesting topic. Could you please point 
> to studies that document this effect, and in particular quantitatively 
> measure its scale (e.g., the fraction of hydrogen being abstracted)? Are 
> there specific types of hydrogen (or particular H sites) that are more prone 
> to this effect, etc.?
>
> Thanks,
> Pavel
>
> On Mon, Jul 20, 2026 at 5:03 AM Rafael Marques 
> <[email protected]> wrote:
>
>> I just would like to add my two cents (again) to the topic when it comes to 
>> the presence of hydrogens. Although we generally assume they are there and 
>> cannot be seen due to their scattering properties, this is not completely 
>> true. Direct radiation damage or induced by water radicals during the x-ray 
>> diffraction experiment may remove them. If one needs to see the hydrogens, 
>> neutron diffraction should be used instead of assuming they are there. At 
>> the end of the day what every structural biologist produces is a model and 
>> one should choose wisely if their model can support or not the hypothesis.
>>
>> 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]> em nome de Gerard Bricogne 
>> <[email protected]>
>> Enviado: domingo, 19 de julho de 2026 22:48
>> Para: [email protected] <[email protected]>
>> Assunto: Re: [ccp4bb] Hydrogen atoms at 3.3 ang
>>
>> Dear Wladek,
>>
>> Doesn't the procedure you advocate lead to the deposition of
>> non-reproducible results? Hydrogens are useful at resolutions at which
>> they cannot be seen because they are involved in contact terms that
>> contribute to producing good geometry by avoiding the kind of clashes
>> that MolProbity hunts down in its quality assessment. If you get that
>> benefit by including hydrogens in HKL-3000 but then omit these in its
>> output file, these refinement results cannot be reproduced by other
>> refinement programs using the information provided by your procedure.
>> They would have to use their own means of reintroducing hydrogens - so
>> why not simply write out "your" hydrogens? It is generally understood
>> that they may not be "experimental" in the sense in which they are in
>> small molecule structures, but that they may have played a significant
>> role in achieving the quality of the final geometry (as judged by
>> MolProbity).
>>
>> Best wishes,
>>
>> Gerard
>>
>> --
>> On Sun, Jul 19, 2026 at 02:31:54PM -0400, Wladek Minor wrote:
>>> Yes, hydrogens are always present, but as opposed to small
>>> molecules, we do not see them in the map.
>>>
>>> Refinement in HKL-3000 includes hydrogens, but the final file does
>>> not include them because they are not experimental. See the paper
>>> that describes state-of-the-art refinement
>>>
>>> Shabalin IG, Porebski PJ, Minor W (2018)
>>> *Refining the macromolecular model - achieving the best agreement
>>> with the data from X-ray diffraction experiment.*
>>> /Crystallogr Rev/24(4): 236-262.[DOI:10.1080/0889311X.2018.1521805]
>>> <https://doi.org/10.1080/0889311X.2018.1521805>[Pub Med ID:
>>> 30416256] 
>>> <https://www.ncbi.nlm.nih.gov/sites/entrez?cmd=search&db=pubmed&term=30416256>[Pub
>>> Med Central ID: PMC6219471] 
>>> <https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6219471/?tool=pmcentrez>
>>>
>>> Wladek
>>>
>>> On 7/17/2026 4:33 PM, Pius Padayatti wrote:
>>> >support overwhelmingly adding hydrogens in all refinements
>>> >why not it is there if one were to be ignore it hydrogens are
>>> >always present
>>> >no matter
>>> >Pius
>>> >_Pius Padayatti_
>>> >
>>> >
>>> >
>>> >
>>> >On Thu, Jul 16, 2026 at 9:09 AM Wladek Minor
>>> ><[email protected]> wrote:
>>> >
>>> > There is another paper about waterless structures.
>>> >
>>> > W.
>>> >
>>> > On 7/16/2026 3:24 AM, Jan Dohnalek wrote:
>>> >> I always have mixed feelings here.
>>> >> Yes, they contribute and "belong" to the model and refinement.
>>> >> On the other hand we (software) does not build all of them,
>>> >> especially on the groups with protonation "freedom".
>>> >> In this way, our refined structures with hydrogens are closer to
>>> >> the reality but they partially lie - as we do not have Hs built
>>> >> in some places where they obviously belong.
>>> >>
>>> >> I would prefer the PDB keeps both - the model with
>>> >> "crystallographic H-set" with all the pluses and minuses it has
>>> >> and then the stripped model - for the reasons above.
>>> >>
>>> >> Jan
>>> >>
>>> >>
>>> >> On Wed, Jul 15, 2026 at 7:57 PM Pavel Afonine
>>> >> <[email protected]> wrote:
>>> >>
>>> >> Hi Flavio,
>>> >>
>>> >> First off, this topic has been discussed a lot on this and
>>> >> other mailing lists, so it might be worthwhile to check the
>>> >> archives. There is even an article on this very topic:
>>> >>
>>> >> https://phenix-online.org/phenixwebsite_static/mainsite/files/newsletter/CCN_2012_01.pdf#page=18
>>> >>
>>> >> From a chemistry and crystallographic software/methods
>>> >> standpoint, I can’t think of a reason why one would even
>>> >> consider not adding H to their models. Hydrogen atoms are
>>> >> present in actual structures; they contribute to the
>>> >> scattering in a measurable way, and they can help improve
>>> >> crystal structure models in many ways that have been
>>> >> discussed before.
>>> >>
>>> >> It’s not about adding or not adding H, it’s about when to add
>>> >> them. It may be helpful (for you and for algorithms) to not
>>> >> include H in the early stages of atomic model
>>> >> building/refinement, but once major errors are finalized,
>>> >> just add H and keep them. For you, because it’s easier to
>>> >> work with a less cluttered model. Same for algorithms:
>>> >> simpler models facilitate convergence and make it easier to
>>> >> overcome energy barriers.
>>> >>
>>> >> And yes, don’t make the terrible mistake of having H
>>> >> throughout the refinement and then stripping them off right
>>> >> before deposition—that will invalidate many of the statistics
>>> >> reported in your model file header.
>>> >>
>>> >> All the best!
>>> >> Pavel
>>> >>
>>> >> On Wed, Jul 15, 2026 at 6:48 AM Flavio Di Pisa
>>> >> <[email protected]> wrote:
>>> >>
>>> >> Dear community,
>>> >>
>>> >> While refining a crystallographic model at 3.3 Å
>>> >> resolution, I noticed an improvement in the geometry
>>> >> statistics after adding hydrogen atoms in riding positions.
>>> >>
>>> >> Would it be inappropriate to keep these hydrogen atoms in
>>> >> the final model to be deposited in the PDB, given the
>>> >> relatively low resolution? Or is it generally preferable
>>> >> to remove them before deposition?
>>> >>
>>> >> Thank you very much in advance for your advice.
>>> >>
>>> >> Best regards,
>>> >>
>>> >> Flavio.
>>> >>
>>> >>
>>> >> ------------------------------------------------------------------------
>>> >>
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>>> >>
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>>> >>
>>> >>
>>> >>
>>> >>
>>> >> -- Jan Dohnalek, Ph.D
>>> >> Institute of Biotechnology
>>> >> Academy of Sciences of the Czech Republic
>>> >> Biocev
>>> >> Prumyslova 595
>>> >> 252 50 Vestec near Prague
>>> >> Czech Republic
>>> >>
>>> >> Tel. +420 325 873 758
>>> >>
>>> >> ------------------------------------------------------------------------
>>> >>
>>> >> To unsubscribe from the CCP4BB list, click the following link:
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>>> >>
>>> > -- Dr. Wladek Minor
>>> > Harrison Distinguished Professor
>>> > University of Virginia
>>> > Department of Molecular Physiology and Biological Physics
>>> > Phone: 434-243-6865
>>> > Fax: 434-243-2981
>>> > https://minorlab.org
>>> > WIKI:https://en.wikipedia.org/wiki/Wladek_Minor
>>> >
>>> >
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>>> >
>>> --
>>> Dr. Wladek Minor
>>> Harrison Distinguished Professor
>>> University of Virginia
>>> Department of Molecular Physiology and Biological Physics
>>> Phone: 434-243-6865
>>> Fax: 434-243-2981
>>> https://minorlab.org
>>> WIKI:https://en.wikipedia.org/wiki/Wladek_Minor
>>>
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