Dear Charles and Xplor-NIH team,

I am a beginner in learning Xplor-NIH and I have a few questions.

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).

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?

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?

Best regards,

Suze Ma
Graduate Student in Chemistry,
Fudan University

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