Biochemistry, Biophysics, and Structural Biology Commons^™

Snake Venom Peptides And Toxin Targeting The Main Protease Of Sars-Cov-2, Breauna Strawder, James Stewart, Mohammad A. Halim

Symposium of Student Scholars

The corona virus began to spread in Wuhan, China which caused it to spread worldwide creating a global pandemic in the beginning of 2020, infecting over 243 million and killing over 4.5 million people worldwide. Significant efforts were made to produce vaccines against the virus, which led the recognition of a few vaccines that has been approved by FDA. These vaccines, Pfizer-BioNTech, Moderna and Johnson & Johnson, which all have efficacy against Covid-19. Despite having vaccines, COVID-19 is still present and infecting millions and killing thousands of people every day. Multiple therapeutic options would allow us to slow down or …

Binding Affinity And Interaction Of Sars-Cov-2 Epitopes With Major Histocompatibility Complex, Sareena Kandavalli, James Stewart, Mohammed A. Halim

Symposium of Student Scholars

SARS CoV-2 has been affecting the world since 2019. It caused 245 million cases of infection and around 5 million deaths worldwide. The most important strategies for the development of vaccines against SARS-CoV-2 are inactivated or weakened virus, replicating or non-replicating viral vector-based approaches, DNA, RNA, virus particle like approaches and epitope-based approaches. The epitope-based approach is rapid, accurate, cost-effective, and reliable against pathogens. By presenting epitopes (antigen peptides) on antigen-presenting cells (APCs), the major histocompatibility complex (MHC), also recognized as the human leukocyte antigen (HLA) system in humans, plays an essential role in triggering T-cell immune responses. The focus …

Cell Penetrating Peptide Inhibiting The Main Protease Of Sars-Cov-2, Adam Ashley, James Stewart, Mohammad Halim

Symposium of Student Scholars

As of October 2021, SARS-CoV-2 has infected over 244 million people and killed about 5 million people. The current FDA approved vaccines are effective; however, they lose their effectiveness after a few months of receiving both doses of the vaccine, and it is recommended to get a booster shot six months after receiving the second dose of the vaccine. Therefore, new highly effective, long lasting antiviral agents and strategies are needed to create an alternative treatment for SARS-CoV-2 and the different variants. Previous studies have shown that cell penetrating peptides (CPPs) have led to greater efficiency of intracellular delivery. However, …

Effects Of Exogenous Application Of Plant Growth Regulators (Snp And Ga3) On Phytoextraction By Switchgrass (Panicum Virgatum L.) Grown In Lead (Pb) Contaminated Soil, Adrianne Beavers, Marina Koether, Thomas C. Mcelroy, Sigurdur Greipsson

Faculty and Research Publications

Soil lead (Pb) contamination is a major environmental and public health risk. Switch-grass (Panicum virgatum), a second-generation biofuel crop, is potentially useful for the long-term phytoremediation and phytoextraction of Pb contaminated soils. We evaluated the efficacy of a coor-dinated foliar application of plant growth regulators and soil fungicide and a chelator in order to optimize phytoextraction. Plants were grown in soil culture under controlled conditions. First, three exogenous nitric oxide (NO) donors were evaluated at multiple concentrations: (1) S-nitroso-N-acetylpenicillamine (SNAP); (2) sodium nitroprusside (SNP); and (3) S-nitrosoglutathione (GSNO). Second, the effect of SNP (0.5 µM) was examined further with the …

The Veiled Lady Fungus, Nick Parbhoo

Symposium of Student Scholars

This semester I studied the Stinkhorn mushroom Phallus Indusiatus. The plan of this research is to develop a protocol for growing this fungus and using it in collaboration with research on it’s web-like properties of the unique veil produced by the fruiting body. This will be distributed to teams of engineers as well as NASA for Dr. Penick’s research. Due to logistics of receiving spores from across the world, we still have not been able to begin growing these mushrooms. However, I have described a protocol that we will follow in order to grow. The protocol contains detailed descriptions …

Conserved Regions Mediate Interactions Between Canonical Nox Domains, Akua Acheampong

Symposium of Student Scholars

NAPDH oxidase (NOXes) are membrane-bound enzymes that generate reactive oxygen species (ROS) that play a role in immune response and signaling. Misregulation of NOXes is implicated in various human pathologies. NOXes contain a catalytic core comprised of a heme-containing transmembrane (TM) domain and a cytoplasmic dehydrogenase (DH) domain that binds FAD and NADPH. Several conserved regions at the interface of the TM and DH domains in eukaryotic NOXes have been suggested to mediate enzyme function and activity. In 2017, researchers successfully purified SpNox, a bona fide NOX homolog from Streptococcus pneumoniae and verified its NOX properties. SpNox’s robust activity in …

Binding Affinity Of Flavins To The Dehydrogenase Domain Of Spnox, Quinesha Williams

Master of Science in Chemical Sciences Theses

NADPH oxidases (NOX’s) are enzymes that catalyze the production of superoxide through single electron transfer. This superoxide production leads to the production of other reactive oxygen species (ROS). ROS affect many metabolic processes throughout the body that can cause several different diseases, making this an ideal target for drug discovery. The general structure of NOX contains a transmembrane (TM) domain and a dehydrogenase (DH) domain connected by a linker. The DH domain contains binding sites for FAD and NADPH/NADH that both participate in the electron transfer necessary for producing superoxide. Structural information of NOX’s is still relatively new to the …

Creating A Protein Chimera To Study Regulation Of Muscle Diversity, Shannon Scarboro

Symposium of Student Scholars

Creating a protein chimera to study regulation of muscle diversity.

Body muscles are made of many individual super-cells, called muscle fibers, that have distinct properties and determine every individual’s strength and endurance. Initially all muscle fibers have identical characteristics, but become differentiated into specific types in adults. The mechanism of such transition is not well understood, despite its obvious importance for shaping human physicality.

Remarkable conservation of the muscle tissue enables us to use fruit flies to study the mechanisms of muscle fiber diversity. We hypothesized that the transcription factor Mef2 acts as a molecular switch that activates structural genes …

Expression And Purification Of Proteins For A Structural Determination: Orf8, Paivy1, Paivy2, Sarah Fashinasi, Avery Moss

Symposium of Student Scholars

Protein purification is a crucial procedure in order to analyze the behavior, structure, and function of a select protein. The Orf8 protein is an increasingly mutable accessory protein found within the SARS-CoV2 virus, however the complex and unpredictable nature of this protein has been met by its inability to be purified thus far. By purifying the model proteins, PaIVY1 and PaIVY2, we believe that we can denote a sound purification method to apply to the Orf8 protein. In our procedure, we transformed BL21(DE3)pLysS cells and inoculated them in LB media. The cells were then placed into a centrifuge to isolate …

Investigating The Sars-Cov-2 Orf 8 Accessory Protein: Expression, Purification, And Structural Determination, Laney Hedgeman, Caroline Salha

Symposium of Student Scholars

Abstract

In order to provide insight into potential therapeutic breakthroughs for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), more research must be done to understand the structure and function of its proteins. The open reading frame 8 (ORF 8) accessory protein is particularly unstable on its own outside of the viral envelope but can be stabilized when bound to the small ubiquitin-related modifier (SUMO) protein. The stable ORF 8-SUMO protein complex can be expressed and purified using familiar techniques and later characterized with nuclear magnetic resonance (NMR) spectroscopy, thus allowing us to gain knowledge about the role it plays …

Bioanalytical Determination Of Glucose Concentration In Sports Drinks Using Uv/Vis Spectroscopy, Laney Hedgeman, Brianna Bond

Symposium of Student Scholars

Abstract

The purpose of our project is to indirectly monitor the enzymatic activity of glucose oxidase as it reacts with the glucose present in sports drinks through the quantitative analysis of the generated ferricyanide. As glucose oxidase catalyzes the oxidation of beta-D-glucose in the presence of oxygen, D-glucono-1,5-lactone is produced along with hydrogen peroxide. The hydrogen peroxide is used in a subsequent reaction with ferrocyanide catalyzed by horse radish peroxidase to produce water and the chromophore ferricyanide. This compound absorbs in the ultraviolet/visible spectrum at 420 nm, which can be quickly measured using Ultraviolet/Visible Spectroscopy. Since the molar ratio of …

Designing A Reactive Warhead To Bind And Inhibit Pseudomonas Aeruginosa’S Periplasmic Protein, Inhibitor Of Vertebrate Lysozyme, Leah Greinke

Master of Science in Chemical Sciences Theses

Pseudomonas aeruginosa is a Gram-negative bacterium commonly found throughout the environment. It is a significant cause of disease and mortality in immunodeficient patients such as those suffering from cystic fibrosis (CF). Due to the emerging antibiotic resistance of P. aeruginosa, it is becoming increasingly more challenging to treat an infection by traditional means. Further complicating treatment, P. aeruginosa secretes a protein known as Inhibitor of Vertebrate Lysozyme (PaIVY) that binds to and inhibits C-type lysozyme, thus preventing the degradation of the bacterium. A reactive chemical warhead was synthesized from a rhenium(I) tricarbonyl derivative inorder to bind to and irreversibly …

Analysis Of Foki Cleavage Resistance Observed In Dna Sequences Generated Via The Combinatorial Selection Method, Repsa, Andre Berry

Master of Science in Chemical Sciences Theses

FokI is a thoroughly investigated and highly utilized restriction endonuclease that recognizes the DNA sequence, 5’-GGATG-3’, and cleaves outside of this site 9 and 13 base-pairs downstream. The shifted cleavage function possessed by this kind of endonuclease is utilized in many applications including the combinatorial selection method, REPSA. FokI employment in the REPSA procedure has demonstrated the tendency to select for an unmodified sequence that possesses the recognition site yet is refractory to cleavage by the enzyme. Sequencing of the cleavage resistant DNA has revealed the inhibitory event to be induced by the presence of an additional inversely oriented recognition …

Probing Interactions Between Canonical Nox Domains, Akua Acheampong

Master of Science in Integrative Biology Theses

NAPDH oxidase enzymes (NOXes) reduce molecular oxygen to superoxide and other ROS. NOXes contain a catalytic core comprising a heme-containing transmembrane (TM) domain and a cytoplasmic dehydrogenase (DH) domain that binds the substrate NADPH and the cofactor. Previously, NOXes were only characterized in eukaryotes, but have recently been identified in prokaryotes, namely bacteria. Due to their constitutive activity and solubility in detergent, bacterial NOXes, such as Streptococcus Pneumoniae NOX, have emerged as a model for studying NOXes. Past research studies in NOXes have identified conserved, putative interacting regions at the interface of the TM and DH domains: the TM B-loop, …