Stacy (Johnson) Duis, Pharm.D.
Education: Bachelor of Science in biology (December 2007)
Doctor of Pharmacy from the University of Kansas
McNair Project: Expression of Recombinant Hepatitis C Glyocoprotein (2006)
Mentor: Rollie Clem, Ph.D.
Hepatitis C Virus (HCV) is a positive, single-stranded RNA virus that infects more than 200 million people worldwide. HCV is a contributory agent of liver cirrhosis, hepatocellular carcinoma, liver failure, and, in 1-5% of all cases, death. Since its discovery in 1989, HCV has been shown to enter cells through an interaction between HCV's envelope glycoprotein, E2, and a cellular receptor, CD81. While this method of entry is widely acknowledged, the role of this interface in the viral life cycle of HCV continues to remain ambiguous. Structural analysis and mutagenesis studies have indicated that the region of CD81 that binds to HCV-E2 is on the solvent-exposed face of helix D, found in CD81's second extracellular loop. Employing this knowledge, imidazole-based compounds were created that mimic the helix D region of CD81, thereby acting as competitive inhibitors of HCV-E2 binding to human CD81. The results of this experiment provided significant evidence that the use of CD81-based mimics can disrupt the interaction between CD81 and HCV-E2, thus inhibiting the entry of HCV into cells.
In the current study, a recombinant baculovirus is being constructed that expresses HCV E2 protein. The E2 protein will be purified for use in further bioassays. These bioassays will be used for supplemental screening and optimization of the CD81 mimics. The purified E2 protein will also be used in characterizing CD81 mutants to determine the effect of the mutations on CD81 binding to E2.