Expression and Characterization of P97

• Luke Coster, Biochemistry, University of Delaware

• Sharon Rozovsky, Department of Chemistry and Biochemistry, University of Delaware

When the cell encounters misfolded/toxic proteins in its endoplasmic reticulum, the cell undergoes the process of endoplasmic reticulum-associated protein degradation (ERAD) to remove such protein. There are multiple pathways for ERAD requiring many different proteins. One important protein of which is P97, a protein that facilitated the ubiquitination of the misfolded proteins, thus tagging the protein for degradation.  To perform p97 task it requires multiple protein partners. One of these partners is Selenoprotein S (SelenoS), yet its role in the function of p97 is still unknown. The Rozovsky Group studies selenoproteins, proteins containing the rare amino acid selenocysteine. One protein of which is the earlier mentioned SelenoS. Therefore, the role of this summer was to purify and characterize p97, so future studies can be done to understand its role in p97’s function. The key method of expressing which was using a recombinant plasmid in E.coli and the key method of characterization being Cryo-EM.