In <3AA2AB65.3E1E52D8 at poczta.onet.pl>, 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?
We can help better if you tell us a little more about your application. Do you
want your protein soluble? Random coil? Disulfides broken (BTW, SDS doesn't
break disulfides)?
In general (N.B. this is grossly oversimplified):
Chaotropes (e.g., urea, guandine, etc.) result in random-coil conformation.
SDS gives a straight rod.
In either case, you need a reducatant to (e.g., 2-mercaptoethanol or DTT) to
break disulfide bonds.
Boiling usually denatures but your protein will most likely be insoluble (think
of a boiled egg white).
Strong acid or base also usually denatures; strong enough acid will also
hydrolyze the peptide bonds.
HTH,
Nick
--
Nick Theodorakis
nicholas_theodorakis at urmc.rochester.edu
