I am studying the function of GLP1r, a GPCR. I want to add 6 myc tags to the C terminal of GLP1r. Can anyone give some information about whether this will affect the function of GLP1r?
I would argue that everything you add/attach has the ability to affect the function...for example adding a ~240 aa large protein (i.e. GFP) to a ~400-500 aa GPCR will probably have some effect..may it be physical in its interactions with other proteins, messengers in your assay system or the direct conformational state of the GPCR itself and therefore its interaction with its ligand. So, you will need some experiments to show that your altered GPCR will act similar as the WT....
The C-Myc-tagged protein contains 11 amino acids, which can still recognize their corresponding antibodies when expressed in different protein frameworks as antigenic epitopes. The c-Myc tag has been successfully used in Western-blot hybridization, immunoprecipitation and flow cytometry, and can be used to detect the expression of recombinant proteins in target cells.
In theory, the larger the molecular weight of the protein tag, the greater the impact on the function of the protein itself, so macromolecular tags are generally only used for detection or protein purification. If the molecular weight of the C-Myc tag is relatively small, the immunogenicity is weak, and generally speaking, it will not affect subsequent research and applications.
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