Fatty acid and retinol binding proteins are a family of proteins found in free living and parasitic nematodes. It is our hypothesis that parasitic nematodes secrete FAR into host tissues, resulting in localized immunosuppression, and hence chronic parsitic infection. Heligmosomoides polygyrus is a mouse model for hookworm that is commonly used in the laboratory for the investigation of the cellular responses to chronic helminth infections. Our laboratory is investigating a protein retrieved from the L4 larval stage of this parasite, a protein believed to be involved in assisting the parasite to form and maintain chronic infections. We have termed this protein Hp‐FAR‐1, for H. polygyrus Fatty Acid and Retinol Binding Protein 1. Creation of mutant FAR proteins that do not bind retinol are of interest for the development of potential vaccine candidates. The purpose of this study was to compare Ce‐FAR‐4, a non‐functional FAR protein from C. elegans, with parasitic FAR proteins. The primary amino acid structures of both parasitic and Ce‐FAR‐4 were analyzed for regions of conservation. Specifically, we target regions that were conserved among the parasitic proteins, including Hp‐FAR‐1, but not in Ce‐FAR‐4; hypothesizing that mutating the conserved regions of Hp‐FAR‐1to the amino acids found in Ce‐FAR‐4 would inhibit retinol binding.

Epstein‐Barr virus (EBV) is a human herpesvirus that causes infectious mononucleosis. EBV is associated with human cancers, such as Burkitt lymphoma and nasopharyngeal carcinoma, due to infection of the B and epithelial cells respectively. The lytic cycle is known as the productive phase of the viral life cycle; in other terms, when the genome of the virus is undergoing transcription and new virions are being produced within a host cell. In the virion, the viral DNA is enclosed by an ordered protein capsid, which is surrounded by a layer of proteins called the tegument. This project studies an Epstein‐Barr virus tegument protein of unknown function. When the gene encoding the uncharacterized tegument protein is knocked out, virion production is not observed.Though important, the structure and function of this lytic tegument protein has yet to be determined. This project has produced pure samples of this tegument protein for study of the basic structural characteristics. The histidine‐tagged protein was produced in E. coli, including C41(DE3) and SHuffle cells. Both strains of cells contain components to promote foreign protein folding, though SHuffle cells contain chaperones specific to disulfide bond formation. When induced in C41(DE3) competent cells, signs of protein degradation were observed on a reducing SDS‐PAGE gel. Successful induction of full‐length protein was observed in SHuffle cells, indicating potential presence of disulfide bonds in the tertiary structure of the protein. The induced protein was purified by Ni2+‐affinity chromatography. Circular dichroism and dynamic light scattering analysis will be conducted to observe structural characteristics in the protein's native state with the goal of determining the structure and predicting a function for this EBV lytic protein.Along with functioning within the lytic cycle of EBV, this tegument protein has potential ties to multiple sclerosis (MS), a central nervous system disease that causes demyelination and degeneration of neurons. From published studies, it has been found that patients with MS have a much greater antigenic response towards one of the tegument proteins. By further studying its structure and function, insight could be gained on not only the protein's effect on the lytic cycle, but also its secondary role it may play in the cause of MS.Support or Funding InformationThis project was supported by UW ‐ La Crosse College of Science and Health, UW ‐ La Crosse Faculty Research Grant, and the Hardy Chan and Sons Undergraduate Research Fellowship.