Hi Charles

Would https://atb.uq.edu.au/ help you here, I am planning to do similar things 
and was planning to use it…

regards
Gary


Dr Gary S Thompson NMR Facility Manager
CCPN CoI & Working Group Member
Wellcome Trust Biomolecular NMR Facility
School of Biosciences,  Division of Natural Sciences
University of Kent,  Canterbury,  Kent,  England,  CT2 7NZ

☎:01227 82 7117
✉️: [email protected]
orchid: orcid.org/0000-0001-9399-7636

On 4 Jun 2024, at 16:28, Charles Schwieters <[email protected]> wrote:

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Hello Suze Ma--


1. I intend to calculate the solution 3D structures of two peptide
macrocycles based on their 1H-1H NOESY data. The first one contains
an ether crosslink (from an exogenous oxygen atom) inserted between
the Trp1-C7 and the Phe3-Cβ while the second one contains a
carbon-carbon crosslink derived from Trp1-C7 and Lys3-Cβ. (for both
structures, see ref: 
https://eur01.safelinks.protection.outlook.com/?url=https%3A%2F%2Fdoi.org%2F10.1021%2Fjacs.3c04355&data=05%7C02%7CG.S.Thompson%40kent.ac.uk%7C004d4f2097b34693a64808dc84ab194b%7C51a9fa563f32449aa7213e3f49aa5e9a%7C0%7C0%7C638531117717912748%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C0%7C%7C%7C&sdata=6NlDB%2FwAT6jU1%2FZjG3b%2BfY9OyFKFbFJJUkrsoq02mxs%3D&reserved=0).
 I am
wondering if there are patches to add these two types of
modifications to the structures. I guess addAtom.py is not suitable
for these cases. In addition, I am not sure how to generate the
parameters (e.g. bond, angle, etc) associated with these
intramolecular crosslinks (I understand that the original protein
topology file, e.g. protein-1.0.top, does not contain such
information).

You will need patches along the lines of those used in
eginput/PSF_generation/gen*CircPep.py in the Xplor-NIH distribution.
I can help with the required patches. But particularly for the ether
case, parameters are required - we might borrow those of anisole, if
you can find the atomic coordinates of that molecule. For the
carbon-carbon crosslink, we can make a decent educated guess.


2. To generate distance restraints, I use sparky to assign the NOESY
spectrum and obtain the intensity of NOE signals, which, upon
calibrated by a known interprotein distance (e.g. Thr-HA to Thr-HN,
2.51 angstrom) and its NOE intensity in my spectrum, only results in
a single list of distances, referred to as d. However, I'm not sure
how to generate the dminus and dplus to define the distance
restraint bounds (d-dminus, d-dplus). I know there are other
methods, such as MARDIGRAS, to generate the bounds. However, I don't
find an available access to these algorithms. Thus, I am asking if
there are other methods to overcome or circumvent the problem in
generating distance restraints bounds for calculations?

Usually, the intensity-derived distances are rather qualitative, so
that distance bounds would be at least +/- 1 Angstrom, so unless you
know otherwise, you might use this value.


3. I am wondering is it possible to only rely on the NOE restraints
(and not dihedral restraints or others) to model the structures of
the small peptide macrocycles in my case? If it is theoretically
feasible, how to calculate only with distance restraints and not
other restraints by using fold.py, anneal.py, etc? Simply by
deleting the contents regarding other restraints while only
maintaining the contents regarding distance restraints?

Of course, more data is better, but distance-only information can be
used by commenting-out or removing sections of the scripts which
involve other restraints.

best regards--
Charles

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