Can anyone suggest a eukaryotic linker for proteins with the property of auto cleavage or protease sensitive and size 3-6 amino acids.
Try this one my dear, I'll be back after checking the literature
Best regards
Article Conserved interdomain linker promotes phase separation of th...
There's the Caspase-3/7 cleavable DEVD sequence. The protease cuts after the second D so you would generally design your linker to be a 7- or 8-mer with the last 3 residues being an inert flexible linker.
TeV (tobacco etch virus) protease is also another good substrate to use if you're looking for something that is non-endogenous to human cells
Protease sensitive: GST-3C protease is my preferred choice with a short recognition sequence. Cleavage is efficient but what's important is removal of the enzyme after cleavage (if you are trying to isolate a recombinant protein?) i.e. binding it to GST beads. This works well in batch purifications. TeV is OK too and is usually His tagged - removal of residual 6xHis-TeV with Nickel resin usually works best with on-column separation. Both resins are quite sticky so you will have to optimise conditions so that your protein does not stick to these and reduce your yield e.g. high NaCl etc
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