Here’s my 2 cents.

If you use a riding hydrogen model (even at low resolution), it affects not 
only the geometry statistics (due to incorporating them for Van der Waals 
clashes), it also improves the agreement of the R-factors between F(obs) and 
F(calc) below about 8 Å. Besides, not including them implies that you don’t 
believe those hydrogen atoms exist in the protein, which we all know isn’t true.

In Phenix, you have the option to explicitly add these hydrogen atoms using the 
ReadySet command. If you use these hydrogens during your refinement cycles and 
for the final coordinates that you use to deposit in the PDB, then include them 
in the PDB deposition. This is no different than thinking you can just delete 
one or two nuisance water molecules in those final coordinates because the PDB 
flagged them as close contacts. The final deposited coordinates should be 
EXACTLY what you used to generate the final set of structure factors from the 
last cycle of refinement.

I wouldn’t worry too much about whether users think that these hydrogen atoms 
are written in stone. Personally I think a lot could be cleared up if the 
output coordinates from such a refinement could say in the CIF file that the 
hydrogen atoms were included as riding atoms, but if someone doesn’t understand 
that you can’t uniquely identify hydrogen atoms at even relatively high 
resolutions in most protein structures, they’re not going to understand what it 
means to use a riding hydrogen model during refinement.

Diana

**************************************************
Diana R. Tomchick
Professor
Departments of Biophysics and Biochemistry
UT Southwestern Medical Center
5323 Harry Hines Blvd.
Rm. ND10.214A
Dallas, TX 75390-8816
[email protected]
(214) 645-6383 (phone)
(214) 645-6353 (fax)



On Jul 15, 2026, at 8:47 AM, Flavio Di Pisa 
<[email protected]> wrote:

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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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