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Addition of fusion protein to C-terminus leads to "escape" from the ER? Is this possible?
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I have a very strange situation regarding a protein construct. I am expressing a fluorescent protein with an ER signal peptide and an NLS right after it. This protein we have shown is secreted. This has been done successfully with both TdTomato and mNeon. There is also a Ty tag at the c-terminus.

Oddly, when we fuse any other protein to the C-terminus (after the ty tag) or even to the n-terminus (after the NLS) the vast majority of the construct then goes to the cell's own nucleus.

How is this possible? I was under the impression that once a protein is sent to he ER it cannot access the cytoplasm. One of the fusion constructs is only around 62kDa (TdTomato is 55kDa) so size doesn't seem to be involved unless there is a very strict cutoff or something.

Thanks in advance, we are scratching our heads on this one.

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PhD|Parasitology, Biochemistry, Cell Biology

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6 years ago