This project falls under the research topics of protein structure, dynamics, function, and biophysical chemistry.The main focus of this project is to elucidate the secondary structure of a viral bacteriophage protein from the ΦKT podoviridae that infects E.coli bacteria. The protein of interest is the gp28 endolysin protein that most likely functions to break the outer membrane of gram negative bacteria during the process of lysis and release. Podoviridae have the ability to infect and kill bacteria without harming surrounding animal or human cells. With antibacterial resistance becoming an increasingly imminent issue in the medical field, having a better understanding of how a virus could kill and inhibit the spread of bacterial growth is extremely useful. The research was conducted by synthesizing the protein of interest using solid state peptide synthesis to introduce cysteine amino acid mutations as a means to attach a spin label probe. Attaching a spin label probe that has an unpaired electron allows dynamic information to be obtained about the amino acid residue position that the spin label is attached to. Once the proteins are synthesized they are integrated into a vesicle membrane that mimics the phospholipid bilayer of a bacterial cell. Techniques such as dynamic light scattering (DLS) and circular dichroism (CD) are used to assess the homogeneous formation of the vesicle, and the helical content of the protein respectively. After these techniques are used, the sample is then analyzed using an electron paramagnetic resonance (EPR) instrument that detects changes and perturbations in a magnetic field caused by the motion and presence of the unpaired electron present in the attached spin label. This project is currently being researched and has not reached a point of major conclusions or findings. We have collected preliminary DLS, CD, and EPR data on a few amino acid positions of interest. The plan going forward is to continue collecting EPR data for more amino acid positions, in addition to employing additional EPR techniques such as double electron- electron resonance, and power saturation. In the fall of 2022 I will be attending Northwestern University’s graduate Interdisciplinary Biological Sciences Program with the intention of receiving my Ph.D. I hope and plan to work on research related to viruses, virus pathways and mechanisms. My work on this project has been a great introduction to viruses and specifically viral lysis proteins.
Authors: Emily Faul, Nancy Rotich, Gary Lorigan
Advisors: Gary Lorigan and Michael Kennedy, Department of Chemistry & Biochemistry
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