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[EMAIL PROTECTED] wrote on 06/01/2006 08:15:06 AM:

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>
> The resolution is 2.5 A.
>
> Indeed, there should be no constraint to tie the ligands together,
> but why do the 2 compounds overlap exactly (in the identical part)
> if I refine only one of them at a time in REFMAC and superimpose the
> structures afterwards ??
> Why do they overlap exactly when I use CNX istead?
>
> Biochemically, I expect them to bind in a similar way, but for sure
> there is no guarantee that they will bind exactly the same way (for
> the identical part).
>
> Regards
> Christian
>

Hi Christian -

Without seeing pictures or rmsd's, I don't know what you mean by "overlap
exactly", but my guess is this: if you only have one molecule to fit
against the data, refmac will give you the same result every time for the
same model - or in your case, the parts of the models that are the same.
However, if you have two superimposed models, refmac now has extra degrees
of freedom to try to fit the data more precisely, and so it tweaks the
models a bit this way and a bit that way so that their combined structure
factors fit the data better than either one could alone.  Now, a good deal
of this "improved" model is probably fitting noise in your data.  But with
good data, the noise level should be low enough that the two copies overlap
reasonably well.

So the question is, how badly are your models failing to overlap?  RMSD >
0.1 A? Or are the lines not right on top of each other in O (which could be
an rmsd as low as ~0.03 A)?  Do the identical parts have significantly
different conformations - torsion angles > 20 degrees different, bonds
flipped, etc.?  Can the different positions of the identical region be
explained by the positions of the varying region?

I'm not sure why CNX gives tighter overlap than refmac does.  Perhaps a
harmonic restraint that you didn't realize you'd assigned?

At the end of the day, I wouldn't worry too much about the variations if
they're small.  Just tell your modelers that this is what happens when you
work with REAL data!

- Matt

--
Matthew Franklin                     phone:(917)606-4116
Senior Scientist, ImClone Systems      fax:(212)645-2054
180 Varick Street, 6th floor
New York, NY 10014


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