disulfide bond and protien folding

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debanjan.pubiot
debanjan.pubiot's picture
disulfide bond and protien folding

Disulfide bond present on the protien help to take the tertiary structure.....is it possible to arrest the protien folding of protien by reagent which will block the formation of disulfide bond and help us to study the transition state during foldimg?

Ivan Delgado
Ivan Delgado's picture
I am not sure if this is what

I am not sure if this is what you are looking for, but in order to break disulfide bonds in proteins all you need to do is add a reducing agent like BMB (beta- or 2-mercaptoethanol) or DTT (dithiothreitol). If you are trying to catch transition states, you could add minimal amounts of this reagents and then increase their concentrations while you monitor the folding state of the proteins. 
If you are looking for something else please let us know.

debanjan.pubiot
debanjan.pubiot's picture
Ivan wrote:

Ivan wrote:

I am not sure if this is what you are looking for, but in order to break disulfide bonds in proteins all you need to do is add a reducing agent like BMB (beta- or 2-mercaptoethanol) or DTT (dithiothreitol). If you are trying to catch transition states, you could add minimal amounts of this reagents and then increase their concentrations while you monitor the folding state of the proteins. 
If you are looking for something else please let us know.

Yup, these are the chemicals which are use to reduce the disulphide bond in porteins.But how far it will be fruitful in studying the folding process by capturing the transition state.How to capture the transition state is there any basic protocol for the study of this process and more over,proteins with higher percentage of disulfide bond can the study to some extent but what abt the proteins without disulfide bond how can be these two be corelated wether they can be or not.... 

debanjan.pubiot
debanjan.pubiot's picture
Ivan wrote:

Ivan wrote:

I am not sure if this is what you are looking for, but in order to break disulfide bonds in proteins all you need to do is add a reducing agent like BMB (beta- or 2-mercaptoethanol) or DTT (dithiothreitol). If you are trying to catch transition states, you could add minimal amounts of this reagents and then increase their concentrations while you monitor the folding state of the proteins. 
If you are looking for something else please let us know.

Yup, these are the chemicals which are use to reduce the disulphide bond in porteins.But how far it will be fruitful in studying the folding process by capturing the transition state.How to capture the transition state is there any basic protocol for the study of this process and more over,proteins with higher percentage of disulfide bond can the study to some extent but what abt the proteins without disulfide bond how can be these two be corelated wether they can be or not.... 

Ivan Delgado
Ivan Delgado's picture
How far it will be fruitful

How far it will be fruitful in studying protein folding? Honestly I do not know how to answer that question. It depends on what you are trying to learn in your experiment. If you are trying to understand how your protein interacts with other molecules, then maybe understanding its folding process may shed some light into this.
As for methods on how to measure this process, there are a number. You can perform circular dichroism spectroscopy for example. Unfortunately I have not done this and I am not aware of any protocols.