I wanted to clone and express few of the nuclear transcription factors (NTFs) in mammalian system. To be more precise, the NTFs are to be secreted in the media from where I have to purify them.
The question of mine is: Whether adding the secretary signal to the system will change its fate? As in it will go through ER complex where post translational modifications occur so will the NTFs will not undergo any such modifications?? Will they behave same as in case of the cytoplasmic expression.
You can express the NTFs with a secretory signal and you may indeed get your protein into the medium, but you should check for modifications later.
You could have a malfolded protein because you get disulfide bridges that would not be formed in the cytosol, and if malfolded the protein might not be secreted. The signal peptide will be cleaved off, but sometimes the exact processing site might not be before the methionine of your protein. If the protein has consensus sites for N-linked glycosylation (NXS or NXT), they can be glycosylated although that is not guaranteed. There are a number of other modifications that you could have, and indeed you need to check for those.
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