Question

Describe in comeplete detail the anfinsen experiment in protein folding.

Answer 1

In 1957 Dr. Christian Anfinsen was the first to put protein renaturation on a quantitative basis with the use of bovine pancreatic RNase. He was awarded the Nobel Prize in Chemistry in 1972. According to Biochemistry by Donald Voet and Judith G. Voet, RNase A, a 124-residue single chain protein, is completely unfolded and its four disulphide bonds reductively cleaved in an 8M urea solution containing 2-mercaptoethanol.

The speaker introduced the experiment by stating the substances 8M urea and β marcaptoethanol. The ribonuclease (the protein used for the experiment) was referred to as the native protein, another term for the primary structure. Mention was also made that the denatured form of the protein forms a random coil with no activity. After the denaturation, dialysis was used to reform the protein to its native form along with the use of air for the oxidation of the disulphide linkage. At this point emphasis was placed upon the fact that all of the information for the protein folding is in the primary structure and that the native form is the most thermodynamically stable.

The importance of the specificity of the disulphide in the correct order was then explained by using visual examples to highlight its importance. Another point made was the fact that some proteins are difficult reform after its denaturation and as a result, they would require other proteins to “chaperon” them to be able to reform their native form.

So the whole experiment is based on the conversation of protein from foldimf to single chain and reverss and check their activities.

I recommend a video by which you can understand clearly -

https://youtu.be/zaaAolMw25A

Thanking you. Hope you get help from the answer.