Paper- Restricted to Campus Access
Metalloproteins use small metal cations or metal containing cofactors to provide specific structural and reactive properties. Their high prevalence makes them of interest to scientists, and one way to study their role in biological systems is through de novo protein design. This method uses novel, lab designed proteins to model natural behaviors, and the Due Ferri single chain (DFsc) protein is one such de novo protein. The DFsc protein is a small, stable, four-helix bundle protein that can bind a wide range of transition metals and is tolerant to site-directed mutagenesis. It was recently shown that the DFsc protein family is capable of hydrolyzing phosphate bonds in nucleic acids and small molecule substrates. The goals of this project are to use site-directed mutagenesis to create four new variants within the DFsc protein family and to continue hydrolase studies with oligomeric DNA and RNA strands to better understand the structure-function relationships of the DFsc fold. The variants created (E11H/H107D- G4, E11H/H107E-G4, E44H/H77D-G4, and E44H/H77E-G4) contain a trans-His conformation in the active site to better model certain natural hydrolases. Sequencing results confirmed successful mutagenesis, and the proteins were produced and purified. RNase and DNase studies are ongoing.
Leach, Danny, "Creation and Characterization of de novo Due Ferri Single Chain Proteins with trans-His Active Site Configurations" (2023). Chemistry Summer Fellows. 48.
Available to Ursinus community only.