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Hi Neil,

It may depend a lot on your expression system, buffers throughout the
purification/crystallization, etc.

My guess it that it looks like N-terminal acetylation. If your protein
is not too big you should be able to see this by ESI-MS.


Cheers,


Miguel

En/na Neil Paterson ha escrit:
> Dear all,
> 
> I'm currently puzzled by some N-terminal density visible in a 1.4
> angstrom map (image <http://www.chem.gla.ac.uk/%7Eneison/blob.png>).  My
> protein has been cleaved with 3C protease, cleavage confirmed by
> SDS-PAGE and failure to rebind a NiNTA column.  3C should cleave
> LEVLFQ/GP yet there appears to be strong density preceding the glycine. 
> The density is not a great fit for an additional residue (although there
> shouldn't be one) and I'm sure my glycine is in the correct place as the
> proline density is very clear. Has anyone seen anything like this before
> or have any suggestions as to what it may be (and how to model it)?
> 
> Thanks,
> Neil

- --
Miguel Ortiz Lombardía
Centro de Investigaciones Oncológicas
C/ Melchor Fernández Almagro, 3
28029 Madrid, Spain
Tel. +34 912 246 900
Fax. +34 912 246 976
e-mail: [EMAIL PROTECTED]
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Et ainsi ne pouvant faire que ce qui est juste fût fort, on a fait que
ce qui est fort fût juste.
                                                Blaise Pascal, Pensées
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