[RASMB] His tagged proteins

[John Philo]

This has been discussed previously in this forum--check the archives.
 
I don't know of any published data off the top of my head, but in several
cases when I was at Amgen we saw dimers and/or higher aggregates in
His-tagged proteins that did not exist for the same protein without the His
tag. We also had cases where the functional properties were significantly
altered by the tag. Hence the use of His tags was largely abandoned there (I
don't know whether that is still true).
 
John Philo

-----Original Message-----
From: rasmb-admin at rasmb-email.bbri.org
[mailto:rasmb-admin at rasmb-email.bbri.org] On Behalf Of John Rodgers
Sent: Tuesday, July 08, 2003 3:11 PM
To: 'RASMB at rasmb-email.bbri.org'
Subject: [RASMB] His tagged proteins



I can imagine problems with performing ultracentrifugation experiments with
His tagged proteins.  It seems that this was also mentioned in the recent
workshop. A set of equilibrium runs were made before the workshop with the
more abundant and available His tagged protein provided by the researcher.
With regard to the protein,  SEC static light scattering experiments were
performed on both native extracted proteins and His tagged protein with
consistent results (dimer-?tetramer in rapid equilibrim.).    The researcher
makes the argument that the His tags are of no significant consequence, but
is open to doing the extra work if a good case can be made for doing so.
Experience, advice and references appreciated.  
Thanks,
John Rodgers  

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