Since both N and C terminals are needed for dimerization they cannot be tagged, I presume. My protein forms N-C dimers that are required for its activity. Any truncation or modification could lead to inactivity.
In theory it should be possible; in practice...
If you have structural information, look for a loop (preferably a disorganized one) and insert your tag there. There's no telling beforehand whether it will work or not for sure, we are not there yet.
OTOH, can't you add a tag to the N or C-termini separated by a flexible spacer?
Hi there,
Inserting a tag in a protein sequence is similar to domain insertion. As long as the sequence is inserted in an solvent-exposed loop of the protein it shouldn't interfere with protein biological activity and the tag should be accessible for whatever you would like to do with it
Hi Subramaniyam, can you introduce a cysteine into the middle of your protein?
That way you can tag it specifically with maleimide fluorophores.
Hello all,
Thank you for your responses. Alejandro, I suspect it will not be possible to use a flexible linker on any terminal since the protein forms an N-C-N-C dimer. Anyways, I shall look into that.
Regards,
Subramaniyam
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