Uma, that is not something that we see regularly in the crystal structures.
But i never used PFPE either. So, you may try PFPE. If this is not PFPE,
then you may read the biology of the protein.

Also I can see green density, but these green blobs are devoid of blue
density. So i will be cautious. May be just add water molecule in each
blob, and re-run refinement. You will see a change in color of the blobs,
that may indicate something.. If red blobs appear around the water
molecules, i will consider it just a noise. Alternatively, you will see
green density connecting the water molecules...





Best wishes

-Z


Zaigham Mahmood Khan, PhD

Icahn School of Medicine at Mount Sinai
Department of Oncological Sciences
1470 Madison Avenue
New York

On Mon, Jul 2, 2018 at 10:23 PM, Uma Gabale <
[email protected]> wrote:

> Dear all,
>
> We came across a blob of unidentified electron density in a shallow cavity of
> a bacterial protein structure (pictures attached). It is surrounded by
> residues Asp, Arg, Gln, His, Glu, Thr, and Trp.
>
> The protein was expressed in *E. coli* BL21(DE3) and purified on Ni-NTA
> followed by gel filtration. The purification buffers included Tris,
> crystallization condition had HEPES and PEG3350; perfluoropolyether was
> used as a cryoprotectant.
>
> We would appreciate any help in identifying it.​
>
> Thanks and regards,
>
> Uma.
> --
> Uma Gabale, PhD
> Research Associate
> Molecular and Cellular Biochemistry
> Indiana University Bloomington
>
>
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