expression of a subunit

Dr Engelbert Buxbaum via methods%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Fri May 29 12:07:27 EST 2009


Am 26.05.2009, 12:50 Uhr, schrieb Azam Rahimpour <rahimpour_a from yahoo.com>:

> I am going to express a mutant subunit of a multimeric complex protein  
> in mammalian cells , this subunit is expected to incorporate in the  
> whole protein complex, I dont know if attachment of small tags like his  
> tag or HA tag would disrupt its structure and prevent its incorporation  
> in a complex. does any one has any experience?

If the 3D structure has been solved (check the PDB), you can look wether  
C- or N-terminus is free (solvent exposed). If that is the case, puting a  
tag there should not overly affect folding and association. The same is  
true for exposed loops, but keep in mind that they may be a binding site  
for regulatory proteins (if that is important to you).

If the structure is not known, then you'll have to use trial and error.


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