Method for studying interaction of a peptide with unknown proteins / peptides
Carol Trim
Wednesday, 13 June 2007 16:41 UTC
I would like to find a cheap and easy method to study the interaction of a known peptide with other unknown peptides / proteins.
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Replies
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Lots of good protocols here
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Hi Carol,
The best method is going to depend on what tools you have available to you and what exactly you want to find out. The size of your peptide will also determine which methods will work for you.
If your peptide is large enough, and you have an antibody specific to it, you might be able to attempt a basic pull-down approach. However, this will not give you genome-wide information on all potential interactors.
For this, you will have to screen an expression library. There are a number of ways to do this and some common approaches include: phage display; yeast-2-hybrid screens – or MAPPIT in mammalian cells; and protein complementation assays (PCAs), including BiFC.
Please note that this is by no means an exhaustive list of methods! And, as each approach has its own advantages and limitations, you will have to do a bit of reading to find out which is best suited to answering your particular question. As a starting point, the following scientists are respected experts in MAPPIT, PCAs and BiFC respectively: Jan Tavernier, Stephen Michnick and Tom Kerppola. These screens can be carried out (relatively cheaply) in cell culture – but may not be appropriate if your peptide is very small as they require the construction of functional fusion proteins.
I hope this gets you started – but let me know if you need information more specific or relevant to your particular experiment and I’ll try to help!
Good luck!
Dot
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