Protein denaturation - (Oct/01/2015 )

Hello,

I hope I am not repeating the topic, if I do please direct me to it

I have a general question and please recommend me if someone knows some good paper or something similar to read on this subject.

I wanna know more about denaturating and renaturating proteins. I know it depends on which protein, its size etc., but I am just asking for some general information at the beginner level (some introduction on methods).

Thank you!

with sds, urea, other salts?

Hi, thank you, but I am asking about something like Anfinsen experiment, I want to renature my protein again, or I want to denature partially because the protein is large...

you can renature from sds with triton or nonidet

you can renature from urea by slowly lowering the concentration (by dialysis)

if you salt out the protein (eg with ammonium sulfate) then you can resuspend with fresh buffer with reduced salt.

i'm not familiar with the "Anfinsen experiment" (at least not by name). if you give details then maybe we can help better.

edit:

i just looked at the "Anfinsen experiment". it's pretty straightforward. denature with mercaptoethanol and urea and renature by diluting urea (and mercaptoethanol) and allowing disulfides to reform. dilution of urea should be performed slowly by stepwise dialysis to ensure proper refolding.

Thank you very much for checking it.

I think I will try Anfinsen's, but also your suggestions!

I thought maybe denaturation-renaturation subject became more advanced since Anfinsen's times and maybe there are some other possibilities I can try, but I guess I was wrong...

http://www.biologicscorp.com/inclusion-bodies-purification-protocol/

In many cases, high-level expression of recombinant proteins leads to the formation of protein aggregation commonly called as inclusion bodies. In order to obtain biologically active and soluble protein in high yield, inclusion bodies must then be solubilized and refolded in vitro.The first step is purification of inclusion bodies. For more information about protein refolding, you can refer to Overview of Protein Refolding.