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what protein-protein interaction can lead to irreversible/covalentbinding

what protein-protein interaction can lead to irreversible/covalentbinding - Protocols and Methods Forum

what protein-protein interaction can lead to irreversible/covalentbinding - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 11-15-2008, 12:50 AM
Zhong Silin
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Default what protein-protein interaction can lead to irreversible/covalentbinding



Hi all

I want to find a method to pulldown a target protein A indirectly. (it justhas to be indirect)

I am thinking: to express protein A with epitope tag x first (A-x), then express a second protein B with another epitope tag y (B-y). Protein B will interact with tag x and Protein A can then be co-purified using the epitope tag y, which is fused to protein B.

For tag x and protein B, I only got 2 pairs of candidates: IgG and protein A/G; N- and C-terminal of the fluorescent protein. Or use the biotin ligaseas protein B and fuse the recognition sequence to protein A as tag x.

The ideal tag x and protein B also have to be small so they wont interfere with the function of protein A. Their interaction has to be strong as well.

I worked on plants and I know our techniques are always a bit behind animalsciences. So you guys must have a better systems to do this. But please keep in mind that this method has to be two-step and indirect!

Thanks in advance
SZ


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