Biochemistry Commons^™

Identification Of Novel Phosphorylation Sites In Sars-Cov-2 Nucleocapsid Protein, Josie Daldegan Rezende

Theses and Dissertations

COVID-19, the 2019 disease outbreak, continues to cause severe mortality worldwide. Fast and reliable tests that identify the SARS-CoV-2 virus, which is responsible for causing the COVID-19 disease, are crucial for controlling the spread of the virus. Antigen tests are ideal for the rapid diagnosis of COVID-19. Current antigen tests for COVID-19 detect the SARS-CoV-2 nucleocapsid (N) protein. The nucleocapsid is the most abundant protein produced by the virus upon infection, which makes it a superior target for detection by an antigen test. Nucleocapsid proteins from several coronaviruses, including SARS, have previously been shown to be modified by posttranslational modification …

Spatially Controlled Monolayers For Electrically Switchable Biomolecule Detection, Eduard Lukhmanov

Theses, Dissertations and Capstones

The development of biosensors that are low-waste, highly stable, and possess an ability to be interchanged between capturing and non-capturing configurations is promising for the biodetector field. Stable packing and attachment of the sensor, the ability to create an interchangeable detecting probe of interest relatively easily, and dynamic control of the probe via a reversible bias can provide for that. With the set goals to control all those properties, DNA-dendron conjugate molecules were designed, synthesized in solution, purified, and utilized to make self-assembled monolayers of single-stranded DNA on gold. To be able to manipulate the conjugates’ sensing qualities in the …

Directed Evolution Studies On A Firefly Luciferase Variant To Improve Signal Intensity For In Vivo Bioluminescence Imaging, Allison Racela

Biochemistry, Cellular and Molecular Biology Honors Papers

In previous research, bioluminescence proteins have been used for in vivo bioluminescence deep tissue imaging. A familiar organism that produces bioluminescent light, is a firefly. Firefly luciferase normally produces yellow-green light (λmax ~ 560 nm) with LH2. There are several reasons for improving bioluminescent light to emit in the near-infrared window (nIR) between 650 nm and 900 nm. The main reasons for improving nIR detectability include that the light transmits more efficiently through mammalian tissues due to less light scattering, nIR light is also less likely to be absorbed by hemoglobin, and water. Overall this provides better resolution for deep …

Characterization Of Cyclopropyl Synthases Involved In The Maturation Of Ribosomally Synthesized And Posttranslationally Modified Peptides, Yi Lien

Electronic Theses and Dissertations

Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a large class of natural products with significant human health implications. RiPPs are synthesized from a genetically encoded precursor peptide that undergoes significant modifications by maturing enzymes, or maturases. Recently, radical-S-adenosylmethionine (rSAM) enzymes have emerged as an important family of RiPP maturases. rSAM enzymes have been shown to install ether, thioether, and carbon-carbon bonds on the precursor peptide. These modifications usually define the backbone structure of the mature RiPP. This thesis describes the characterization of a novel RiPP modification catalyzed by the radical S-adenosylmethionine enzyme TigE. TigE belongs to the TIG biosynthetic …

Inhibition Of De Novo And The Prion-Like Spread Of Amyloidogenesis Using In Vitro And In Vivo Disease Models, Johnson Anazoba Joseph

Electronic Theses and Dissertations

The aberrant fibrous, extracellular, and intracellular proteinaceous deposits in cells, organs and tissues are referred to as amyloids. These deposits are dominated by β-sheet structures that have been implicated in several neurodegenerative diseases and cancer. In this work, the types of amyloidosis studied include Parkinson’s disease (PD) using UA196 and NL5901 strains of Caenorhabditis elegans (C. elegans), Alzheimer’s disease (AD) using GMC101 strain of C. elegans, and cancer-associated mutant p53 aggregation in MIA PaCa-2 mutant cells. Several molecules including SK-129, NS132, NS163, bexarotene, a polyphenol (-)-epi-gallocatechine gallate (EGCG), ADH40, RD148, and RD242 were screened in vitro and in …

Lipophilic Probes For Cellular Ethylene Detection, Morgan R. Brown

Electronic Theses and Dissertations

The structure of ethylene is simple, yet its biological effects are significant. When considering its role in biology it is almost exclusively regarded as a plant hormone. Research on ethylene from plants was progressed by several advancements in analytical instrumentation, from its discovery to elucidation of its signaling pathway. There is currently limited understanding of ethylene’s role in mammals, but evidence suggests that it may be a biomarker for oxidative stress! Additional tools and technology are crucial to study this surprising and important signaling role in mammals. Our group has developed molecular ethylene probes as a strategy to detect ethylene …

Cysteine-Tyrosine Crosslink Synthesis In Bf4112 From B. Fragilis, Alex Van Uffelen, Sophie Hruska, David Benson

Summer Research

Redox-active crosslinked amino acids are a class of post-translational modifications that extend protein chemistry. These cofactors occur in fifteen known proteins with high resolution Xray crystallography as the primary method for confirmation. One of those proteins is cytochrome c oxidase (complex IV), where HisTyr (figure 1) is responsible for the high-valent oxidative intermediate in O2 reduction to 2H2O. Cys-Tyr was the first of these cofactors to be identified and is characterized in galactose oxidase (coordinated to a Cu ion) and cysteine dioxygenase (distal to a mononuclear non-heme iron ion).

The Effects Of Neuronal Growth Factors On Lps-Activated Schwann Cells, Caitlyn E. Henry, Peyton Kimmel, Angela Asirvatham Ph.D.

Student Research Poster Presentations 2022

Schwann cells (SCs) are the principal support cells of neurons in the peripheral nervous system, that both myelinate axons for the rapid conduction of electrical impulses as well as assist in axonal repair during nerve injury. During nerve injury, SCs secrete tumor necrosis factor alpha (TNF-α)^1,5,6 and other proinflammatory mediators^1,6, attracting macrophages to the site of injury to induce inflammation and clear myelin debris.^1,6 Once the debris is cleared, the neuron stimulates SC proliferation by secreting neuronal mitogens, such as heregulin^3,4, and an unknown factor that activates the cAMP pathway³, an important …

Simulation Of An Inflammatory Model Using Schwann Cells, Caitlyn Henry, Peyton Kimmel, Angela Asirvatham

Student Research Poster Presentations 2022

During peripheral nerve injury, the myelin surrounding the neuronal axons is damaged, initiating an inflammatory response to remove myelin debris. Once myelin debris is cleared, Schwann cells acquire a proliferating phenotype which allows them to grow and divide so that remyelination can occur. The neuron stimulates Schwann cell division by secreting growth factors, like heregulin, and an unknown growth factor that activates the cAMP pathway. Although the role of cAMP in axonal regeneration is well-known, not much has been explored about its function in Schwann cells during nerve injury and inflammation. To simulate an inflammatory environment, the S16 Schwann cell …

Using Circular Dichroism Spectroscopy To Study Secondary Structure Of Horseradish Peroxidase: A Laboratory Experiment In Biochemistry, Shemaiah Kamran, Sarah E. Bultje, Kumar Sinniah

Summer Research

• We present a laboratory experiment in biochemistry to study the Far-UV Visible secondary structure of a protein (horseradish peroxidase, HRP) that can be adapted for a single-session laboratory experiment or a multiple-session laboratory project.
• A Circular Dichroism (CD) spectrometer provides structural information of proteins as a function of environmental changes such as pH and temperature. We present secondary structural information for the protein, horseradish peroxidase, at pH 3 and pH 7 in the temperature range 30-80° C in phosphate buffered solution (PBS).
• HRP has many applications in biochemistry and biotechnology. It is frequently used as a reporter enzyme in diagnostics …

Affinity Of Cadmium Versus Zinc For The Carbonic Anhydrase Active Site, Rachel Phan

Honors Scholars Collaborative Projects

Cadmium is a heavy metal found in tobacco plants, nickel-cadmium batteries, and as a byproduct of metal mining. The similarities between the properties of zinc ions and cadmium ions in proteins due to their 2+ charges may cause displacement of zinc by cadmium if encountered and has been shown to do so in several transcription factors. Cadmium’s ability to replace zinc- a metal found in around 3,000 human proteins- is a cause for concern, especially due to its toxicity, long half-life in the body, and larger size.

Carbonic anhydrase is an important metalloprotein protein that relies on a central zinc(II) …

The Role Of Dichloro(Ethylenediamine) Platinum Ii In Altering Cellular Iron And Copper Transport, Megan Hall

Mahurin Honors College Capstone Experience/Thesis Projects

Approximately 50% of all chemotherapy cancer patients receive a platinum chemotherapeutic during their treatment. Although these drugs are widely utilized, little is known about the cellular transport of platinum drugs. It is hypothesized that platinum utilizes transporters for other metals to enter the cell. Structural differences between compounds are predicted to affect the compounds’ ability to exploit different transporters. Also, different cell types express different transporters. Therefore, compound structure is expected to influence transport of platinum compounds differently in different cell types. This study investigated the role of the platinum drug cisplatin and platinum compound Pt(en)Cl₂ in altering iron …

Investigating Spatiotemporal Kinetics, Dynamics, And Mechanism Of Exosome Release, Anarkali Mahmood

Electronic Theses and Dissertations

Exosomes are small lipid-based vesicles that can carry biomolecules from one cell to another. While exosomes are crucial to maintain homeostasis in healthy cells, they are exploited by unhealthy cells to aid disease progression. Exosomes likely facilitate disease progression via the transfer of disease-causing biomolecules from unhealthy to healthy cells. Exosomes are generated in Multivesicular endosomes (MVEs) and are then secreted into the extracellular space to travel to other cells. Despite being a crucial step, very little is known about exosomes release mechanism and dynamics. To further our understanding of exosomes, specifically their secretion, my work has focused on investigating …

Radiation Exposure Determination In A Secure, Cloudbased Online Environment, Ben C. Shirley, Eliseos J. Mucaki, Joan H.M. Knoll, Peter Rogan

Biochemistry Publications

Rapid sample processing and interpretation of estimated exposures will be critical for triaging exposed individuals after a major radiation incident. The dicentric chromosome (DC) assay assesses absorbed radiation using metaphase cells from blood. The Automated Dicentric Chromosome Identifier and Dose Estimator System (ADCI) identifies DCs and determines radiation doses. This study aimed to broaden accessibility and speed of this system, while protecting data and software integrity. ADCI Online is a secure web-streaming platform accessible worldwide from local servers. Cloud-based systems containing data and software are separated until they are linked for radiation exposure estimation. Dose estimates are identical to ADCI …

Kinase-Catalyzed Labeling To Identify Kinase-Substrate Pairs Using Γ-Phosphate Modified Atp Analogs, Rachel Beltman

Wayne State University Dissertations

Post-translational modifications (PTMs) are responsible for a variety of cellular processes. One such PTM is protein phosphorylation, which is catalyzed by kinases. Kinase enzymes play important roles in cellular signaling pathways, but dysregulation of kinase-mediated events results in the formation of diseases, which make kinases favorable drug targets. To uncover the role kinases play in the development of diseases, kinase-mediated cellular events need to be better understood. The current gap in the field is the lack of tools available to identify the kinase that is responsible for specific phosphorylation events within the cell. To improve the gap in the field, …

Metabolic Synergy Between Human Symbionts Bacteroides And Methanobrevibacter, Jennie L. Catlett, Sean Carr, Mikaela Cashman, Megan D. Smith, Mary Walter, Zahmeeth Sakkaff, Christine A. Kelley, Massimiliano Pierobon, Myra B. Cohen, Nicole R. Buan

Department of Biochemistry: Faculty Publications

ABSTRACT Trophic interactions between microbes are postulated to determine whether a host microbiome is healthy or causes predisposition to disease. Two abundant taxa, the Gram-negative heterotrophic bacterium Bacteroides thetaiotaomicron and the methanogenic archaeon Methanobrevibacter smithii, are proposed to have a synergistic metabolic relationship. Both organisms play vital roles in human gut health; B. thetaiotaomicron assists the host by fermenting dietary polysaccharides, whereas M. smithii consumes end-stage fermentation products and is hypothesized to relieve feedback inhibition of upstream microbes such as B. thetaiotaomicron. To study their metabolic interactions, we defined and optimized a coculture system and used software testing …

Arginine Catabolism And Polyamine Biosynthesis Pathway Disparities Within Francisella Tularensis Subpopulations, Yinshi Yue, Bhanwar Lal Puniya, Tomáš Helikar, Benjamin Girardo, Steven H. Hinrichs, Marilyn A. Larson

Department of Biochemistry: Faculty Publications

Francisella tularensis is a highly infectious zoonotic pathogen with as few as 10 organisms causing tularemia, a disease that is fatal if untreated. Although F. tularensis subspecies tularensis (type A) and subspecies holarctica (type B) share over 99.5% average nucleotide identity, notable differences exist in genomic organization and pathogenicity. The type A clade has been further divided into subtypes A.I and A.II, with A.I strains being recognized as some of the most virulent bacterial pathogens known. In this study, we report on major disparities that exist between the F. tularensis subpopulations in arginine catabolism and subsequent polyamine biosynthesis. The genes …

Principles Of General Chemistry, Jose Cobo

Open Educational Resources

No abstract provided.

Investigation Of Catalysis Of Nitration By Cytochrome P450s, Lannika Johnson

Honors Undergraduate Theses

TxtE is a protein related to cytochrome P450 enzymes, which catalyze a number of reactions that typically involve oxygen and not nitrogen. It has been discovered that TxtE can nitrate tryptophan through an unusual reaction in which it uses nitric oxide (NO) as a nitrogen donor to install the nitro group despite NO typically being considered toxic to bacteria. This project will determine if all cytochromes P450 can catalyze nitration as long as they are given NO. This will have an impact on understanding drug delivery and metabolism for which nitration is important.

An Investigation Into The Mechanism Of Proteasome Dysfunction In Neurodegenerative Disease And The Biological Impact Of Proteasome Hyperactivation In C. Elegans, Raymond T. Anderson

Graduate Theses, Dissertations, and Problem Reports (ETD)

Aging is an inevitable process that occurs as humans grow older. It is characterized by the chronological accumulation of cellular damage over time leading to functional decline as an organism grows older. Several processes are thought to contribute to the aging phenomenon, but one of the most prolific of these is the disruption of protein homeostasis (proteostasis). The collapse of proteostasis can lead to accelerated aging and the development of age-related diseases including devastating neurodegenerative diseases (NDs) like Alzheimer and Parkinson disease. Virtually all NDs are characterized by the buildup of proteins in and around neurons resulting in neuronal death …