Link to bioRxiv paper:

http://biorxiv.org/cgi/content/short/2020.09.22.309005v1?rss=1

Authors: Bromberg, R., Guo, Y., Plymire, D. A., Emde, T., Puzio, M., Borek, D., Otwinowski, Z.

Abstract:

The His-tag is a widely used affinity tag that facilitates purification of recombinant proteins for functional and structural studies. We show here that His-tag presence affects how coproheme decarboxylase interacts with the water-air interface during grid preparation for cryoEM. Depending on His-tag presence or absence, we observe significant changes in patterns of preferred orientation. The analysis of particle orientations suggests that His-tag presence can mask the hydrophobic patches on a protein's surface that mediate the interactions with the water-air interface, while the hydrophobic linker between a His-tag and the coding sequence of the protein may enhance other interactions with water-air interface. Our observations suggest that tagging, including rational design of the linkers between an affinity tag and a protein of interest, offer a promising approach to modulating interactions with the water-air interface.

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