Thanks Eric!

I was about to say that (plus a blank .frcmod file) is all you'd need for a case like this (where the residue:residue linkage is a simple, well-parameterised amide bond) - but that would run into the issue that the standard AMBER forcefield doesn't include any parameters for (standard atom):(GAFF atom) combinations so it'd fail on the protein backbone. I'm totally swamped at the moment so won't be able to experiment myself for a while, but Yang: if you want to try it out what should work is to use your favourite text editor to manually replace all the GAFF atom types with the corresponding protein ones (you can find a table of GAFF atom types at https://emleddin.github.io/comp-chem-website/AMBERguide-AMBER-atom-types.html and standard types at https://www.uoxray.uoregon.edu/local/manuals/biosym/discovery/General/Forcefields/AMBER.html - use the all-atom carbons in the latter).

To convert to the format needed by ISOLDE, you'll need to first save each residue in isolation (which - Eric, correct me if I'm wrong - will require you to first split the structure using e.g. "split #1 atoms :XXY atoms :XXZ", followed by "save XXY.mol2 #1.1 gaff t; save XXZ.mol2 #1.2 gaff t") then create a couple of dummy .frcmod files (XXY.frcmod and XXZ.frcmod) in the same directory as the .mol2 files. I'm not sure of the minimal contents of a .frcmod, but you could start with the attached and see how that goes. To actually make the conversion, start ISOLDE, then, in the top ChimeraX menu, choose ISOLDE/Prep/Convert AMBER Files. If all goes well, that should create a .xml file for each .mol2/.frcmod pair it finds in the current working directory. In terms of converting over from GAFF to standard AMBER atom names, you can choose to do that either in the .mol2 or in the .xml files (personal preference, but I'd probably choose the latter since it doesn't have the fiddly layout requirements of .mol2).

Good luck... while I'd love to provide something much neater than this, at the moment I truly don't know when I'll find the necessary time.

Best,
Tristan

On Wed, Apr 9, 2025 at 8:04 PM Eric Pettersen via ChimeraX-users <chimerax-users@cgl.ucsf.edu> wrote:
Hi Yang,
I'm not certain what parameters exactly you need, but you can get ChimeraX to give you partial charges and GAFF atom types for the isopeptide-bonded residues.  First you would give the residues non-standard names so that ChimeraX doesn't just look up the parameters in a table, e.g.:

setattr residue1 r name XXY
setattr residue2 r name XXZ

The you just run Add Charge on your structure.  You can write out the GAFF types and partial charges using Mol2 format, e.g.:

save ~/parameterized.mol2 gaff true

--Eric

On Apr 9, 2025, at 7:19 AM, Yang Lee via ChimeraX-users <chimerax-users@cgl.ucsf.edu> wrote:

Hi Tristan and Eric,

From hopelessness, I find myself newly heartened!

I tried but failed to find mention of this isopeptide bond treatment in the original Chimera mailing list archives. Is the procedure mentioned here already part of the Add Charge menu function or a lower level function I can adapt? Also, is it something that can be easily jerry-rigged into ISOLDE's minimisation perhaps with a code change?

Cheers,
Yang
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