N-terminal degradation of Recombinant protein
Dr Engelbert Buxbaum
engelbert_buxbaum at hotmail.com
Wed Feb 15 07:51:49 EST 2006
Russell Wrobel wrote:
> I'm part of a structural genomics group and we do high though put
> protein production in E. coli. N-term. tags can usually be cleaved off
> after protein purification with site specific proteases. In our program
> we have had good luck expressing things with an N-term maltose binding
> protein (MBP) tag. After purification we remove the tag using TEV
> protease. There are several commercially available expression systems
> that rely on the cleavablity of solubility tags.
MBP is a wonderful aid for expressing proteins in bacteria, it keeps
them stable and soluble. However, in many cases affinity purification on
sugar-columns does not work. I had MBP-cyclins once which I had to
purify by conventional means (AS precipitation followed by SEC and IEC)
because of this problem.
I recently saw a paper (Protein Science, IIRC) where authors suggested
an additional His-tag N-terminal of MBP to get the stabilisation of MBP
and the easy purification of Oligo-His.
More information about the Methods