Purify a His Tag protein for Crystallization

D.K. dk at no.email.thankstospam.net
Mon Jan 13 22:19:26 EST 2003


In article <Pine.SOL.4.33.0301131030090.19612-100000 at mcmail.cis.mcmaster.ca>, Kyle Legate <legatek at mcmail.cis.mcmaster.ca> wrote:
>On 13 Jan 2003 mrsankar_in at yahoo.co.uk wrote:
>
>> I have a His Tag protein overexpressed in E.coli and it does not go into
> inclusion bodies. I want to crystallize the protein for which i need it in a
> very high concentration and in very pure form. What way i can purify the
> protein in Ni Column? Denaturing or native conditions?

This is all protein-dependent. Try and see what works best for your
protein.

>What info do you hope to gain from a crystal of denatured protein?

If a protein is partially folded/denatured, it can give a nice idea 
about folding pathway(s). Problem is, such things seem to never 
crystallize. I know I tried :-) 

But I think the original poster simply wants to purify a [folded] 
protein and does not know how to proceed. 

DK



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