Greetings to all. Currently, I have successfully purified the GST-tagged fusion protein. The protein degrades as soon as I remove the tag. It is an inherently disordered membrane protein. Please let me know if there are any suggestions for using this protein with a tag for Cryoem. The combined molecular weight of the protein and tag is 84 kDa. Mw of protein is 43kDa.
Thanks,
Of course, you can determine the structure of the fusion protein. Intrinsically disordered proteins (IDP) often adopt a structure when bound to another protein, and that structure is specific for that interaction. When bound to another protein, the structure may be completely different. In fig. 10.1 of my "Fundamentals of protein structure and function" I have shown that using the example of p53, whose C-terminus may be a coil or an alpha-helix, depending on the partner. Thus, with IDP it is difficult to speak of "native" structure, there may be many of them. Think of what the physiological binding partner(s) of your protein is/are, and how you could stabilise your protein by binding to those.
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