Protein denaturation.

Scott Coutts scott.coutts at med.monash.edu.au
Mon Mar 5 04:20:13 EST 2001


Why do you need to denature your protein? If you tell us what your
application is, it will be easier to give a specific answer. When you
say 'destroy', I assume you want to 'denature' your protein.

The processes you mention (sanger and edman) are usually for sequencing
your protein (finding the amino acid sequence). They remove one amino
acid at a time.

If you want to 'denature' your protein (ie disrupt it's tertiary and
quarternary structure), you can incubate it in 8M urea or 6M guanidinium
hydrochloride (denaturants). Simply suspend your protein in some type of
buffer (HEPES, TE, PBS, whatever) containing the deaturant and incubate
for an hour or so (depends on the protein) at room temperature with
shaking. Some proteins are insoluble in 8M urea, so Gdn.HCl may be
better. A more aggressive way is to simply add SDS to 1% and boil for 10
minutes.

Search the net for 'protein solubilisation protcol' and you'll find
detailed descriptions of how to denature proteins. 


"andrzej f. lyskowski" wrote:
> 
> Hi,
> 
> I know that what I'm going to ask you will be a simple problem, but this
> is my last resort.
> I was ask to dig some information about protein denaturation. Not any
> specific protein and not necessary danaturation in controlled way.
> Simply saying how to destroy protein. I found almost all existing
> information on Sanger and Edman degradation and then that you can do
> hyrolysis or that SDS destroys sulfide briges but not DETAILS.
> 
> Can anyone point me to the place on the net or send me some information
> concerning that subject?
> 
> I am deeply thankfull for any help you are willing to give.
> Andrzej
> --
> 
>  ANY SUFFICIENTLY ADVANCED TECHNOLOGY
>  IS
>  INDISTINGUISHABLE FROM MAGIC.
>         ARTHUR C. CLARK

-- 
Scott J. Coutts                 
------------------------------------------------------------
Bacterial Pathogenesis Research Group
Monash University, Australia
Phone: +61 3 9905 4838       
Email: scott.coutts at med.monash.edu.au 
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