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://doi.org/10.1021/jacs.3c04355). 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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