Hi everyone,
here's my problem, I'm working on the potential role in carcinogenesis of a small protein of 50 aa (E6*I from Human Papillomavirus type 16). Many cellular effects caused by this protein have been described but all studies used tagged protein, never the native one since it's undetectable by Western (unless you work with a surexpression model treated with MG132...)
So I was wondering if you know some publications describing how a tag (HA-, Flag-, myc-, ...) could affect protein properties such as stability, structure, localisation.
Thanks for the help you can provide !
I assume that there is no marked influence of the small tag in definitely in delivery (since the signal in the peptide itself). The commercial tags are optimized for varying sizes of peptides for keeping the the structure and stability from changing the original properties. It may depends on the kinds of study you are doing. I have worked on 17~30kDa with Myc tag and GFP, RFP without any hinderance within the cells. People are discussing structure and chemical modification when they isolated in man-made condition for the study. In cell state of mixed soup in the salt solution of cytoplasm, a tagged golgi protein and a nuclear protein with larger tag did report about the ther behavior in live cells. It is matter of the extent of overexpression. The cells tend to show aggregates within certain vesicles only in a very small number of highly expressed cells. So I think it would have no major huddle for the study of function of the peptide within the cells unless you are looking of chemical structure.
For the interest of more specific matters.
Mol. BioSyst., 2016, 12, 1731-1745
Appl Microbiol Biotechnol (2003) 60:523–533
https://www.researchgate.net/post/Will_His-tag_affect_the_structure_or_activity_of_a_protein
The short answers is- it depends on several factors including the tag type and question one is asking. For your indicated purpose, the Myc-tag may be appropriate to have.
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