Biochemistry, Biophysics, and Structural Biology Commons^™

Protein-Metal Interactions Of A-Synuclein And Bioactive Transition Metals Via Molecular Dynamics, Gonzalo R. Vazquez

Theses and Dissertations

Parkinson’s Disease (PD) is a neurodegenerative movement disorder which has been found to target the dopaminergic neurons of the substantia nigra. It is characterized by the formation of masses known as Lewy Bodies, formed by aggregates of the intrinsically disordered protein (IDP) -synuclein (asyn). Research suggests that alpha synuclein can undergo aggregation in the presence of heavy metals, which may accumulate in the body through a variety of mechanisms. To study how metals may play a role in wildtype (WT) asyn aggregation, MD simulation of the protein was performed in the presence of bioactive transition metals including Fe2+, Fe3+, Cu+, …

Biological And Chemical Evaluation Of Amide-Based Cjoc42 Derivatives As Potential Gankyrin-Inhibitors For The Treatment Of Breast, Lung, And Liver Cancer, Benita Britton-George

Theses and Dissertations

Gankyrin is an ankyrin-repeat protein overexpressed in certain cancers and is responsible for cell growth and proliferation. Gankyrin is important to the development of many types of cancer, including breast, lung, and liver cancer. Consequently, gankyrin and its various protein-protein interactions, are prospective therapeutic targets for inhibiting the progression of certain cancers. In 2016, the first small molecule inhibitor of gankyrin, cjoc42, was discovered and subsequently demonstrated modest antiproliferative activity against liver cancer cells. Previous work from our lab demonstrated that replacing the sulfonate ester of cjoc42 with an amide group significantly improved gankyrin binding while enhancing antiproliferative activity against …

Expression And Function Of Floral Symmetry Genes In The Regulation Of Flower Shape In Fedia Graciliflora, Theresa A. Mustacchio

Theses and Dissertations

Shifts between radial symmetry (actinomorphy) and bilateral symmetry (zygomorphy) in flowers have occurred multiple times independently within angiosperms (flowering plants). Evidence across angiosperms indicate the transcription factors RADIALIS (RAD), DIVARICATA (DIV), and CYCLOIDEA (CYC) play a major role in regulating floral symmetry and morphology. The CYC-like genes have been shown to shift their expression dorsally independently in nearly all shifts to bilaterally symmetrical flowers examined. Three core eudicot clades of CYC-like genes have been identified: CYC1, CYC2 and CYC3, with only CYC2 genes being shown to have a role in floral symmetry. We use the non-model plant, Fedia graciliflora (Caprifoliaceae) …

Functional Analysis Of Ligand Binding Domain 2 Of Outer Surface Protein C Of Borreliella Burgdorferi, Gavin Z. Chambers

Theses and Dissertations

Lyme disease, caused by the spirochete Borreliella burgdorferi, is the most common vector-borne infection in North America. A critical determinant of early mammalian infection is outer surface protein C (OspC), a highly variable lipoprotein that is indispensable for spirochete survival but whose precise function remains unresolved. OspC has been proposed to mediate diverse roles, including complement evasion, adhesion, tissue dissemination, and antiphagocytic activity, supporting the view that it is a multifunctional virulence factor. Structural studies have identified two putative ligand-binding domains (LBD1 and LBD2), with LBD2 forming a solvent-exposed crown at the membrane-distal end of the OspC dimer.

To …

Targeting Als: A Multi-Cellular Study On The Potential Therapeutic Role Of Azole-Based Compounds, Alisha R. Kennedy

Theses and Dissertations

Amyotrophic Lateral Sclerosis (ALS), also known as Lou Gehrig’s disease, is a progressive neurodegenerative disorder that leads to the degeneration of motor neurons, which control voluntary muscle movements such as eating, speaking, and breathing. As these neurons are damaged, voluntary movement ceases, leading to a fatal outcome. The survival time of ALS patients varies, ranging from a few months to up to 10 years. Chapter 1 of this thesis provides an overview of ALS, highlighting the role of transactive response DNA-binding protein 43 (TDP-43), a key feature in ALS and other neurodegenerative diseases. The aggregation of TDP-43, particularly due to …

Site-Specific Modification Of Antibodies Utilizing Microbial Transglutaminase For Improved Immunoassay Performance, Emily Beitello

Theses and Dissertations

Immobilization strategies used for fixing antibodies to solid phase surfaces significantly contribute to the antigen capture capacity and influences the performance and sensitivity of immunoassays. Traditional methodologies result in the random orientation and/or the weak attachment of immobilized antibodies. Different formats of immunoassays scope in applicability to many fields of research and everyday use. Improving the efficiency of immobilizing antibodies in immunoassays is a beneficial endeavor to increase assay sensitivity. Here, we investigated the use of enzyme-mediated conjugation of antibodies to facilitate a highly oriented immobilization. Microbial transglutaminase (mTG) catalyzes the covalent bond formation between a plethora of primary amine …

Caspases Protect Against Hyperglycemia Induced Necroptosis-Like Cell Death In Brain Endothelial Cells, Aleena S. Mathew

Theses and Dissertations

Acute diabetes characterized by high spikes in blood glucose levels in association with severe hyperglycemia can lead to an increased risk of ischemic stroke in the brain. To better understand high blood glucose levels, hyperglycemia, in the brain we explored its effects on endothelial cells. Endothelial cells line the inner walls of blood vessels and are directly impacted in hyperglycemia. Brain endothelial cells are a major component of the blood brain barrier (BBB) which tightly regulates the brain’s microenvironment. The type of cell death such as apoptosis, caspase dependent, or necroptosis, caspase independent, can exacerbate diabetes associated complications. There is …

The Bacteriostatic, Regenerative, And Immunomodulatory Properties Of Extracellular Matrix Particles For Lung Injury, Keera P. Rhoads

Theses and Dissertations

Acute respiratory distress syndrome (ARDS) is a prevalent, life-threatening lung condition, affecting nearly 200,000 Americans annually, with a 40% international mortality rate. There is no cure for ARDS, and current pharmacological treatments have limited effectiveness. Symptoms can be mitigated with mechanical ventilation, though this often leads to ventilator-induced lung injuries (VILI) and puts critically ill patients at risk of infections, including ventilator-associated pneumonia (VAP). A promising therapeutic is the extracellular matrix (ECM), a complex network of structural proteins and bioactive molecules that has been shown to have anti-inflammatory properties and prevent fibrosis. We aim to utilize the regenerative and immunomodulatory …

Unleashing Serine Palmitoyltransferase In Test Tubes And Mice, Matthew W. Peart

Theses and Dissertations

Canonically known both for structural contributions to lipid bilayers and roles in cell signaling, the sphingolipids comprise a dynamic, multifaceted class of molecules which are studied to understand homeostatic cell biology as well as pathophysiology. Sphingolipids contribute to tissue-specific structures such as myelin, a multilayer membrane that allows for rapid conduction of action potentials along neuronal axons. All sphingolipids are downstream products of the rate-limiting and initiating enzyme in the de novo sphingolipid synthesis pathway, Serine palmitoyltransferase (SPT). Demonstrative of its status as the enzyme that catalyzes a crucial step in a biochemical pathway, SPT activity is strictly regulated. This …

Development Of Novel Nanocarriers With Multi Modal Action Against Lung Cancer, Gayathri K

Theses and Dissertations

Lung cancer ranks among the top causes of cancer-related deaths worldwide. The deteriorating environment and unhealthy lifestyle in the modern era has further contributed to the exponential increase in lung cancer incidence across the globe. Lung cancers fall under two major types based on the location of the tumour in the lung namely, small-cell lung cancer (SCLC) which is more aggressive, and non-small-cell lung cancer (NSCLC) which is more prevalent. Late diagnosis has been a major cause for poor prognosis as the cancer is detected only in an advanced stage.

Conventional therapies like chemotherapy, surgery and radiotherapy against lung cancer …

Ecological And Pathological Applications Of The Heat Shock Response, Taylor Rae Stanley

Theses and Dissertations

The heat shock response (HSR) is a cytoprotective stress response pathway that regulates cellular proteostasis. The HSR is an evolutionarily conserved pathway that is essential for normal cellular functioning. Here, we explore the broad ecological and pathological impacts of the HSR. In an ecological context, we perform gene level analysis of the transcriptomes of two closely related sunfish. We found that the more invasive bluegill sunfish has gene expansions in two HSR gene families, the HSP70 family and the HSP90 family compared to the redear sunfish. These gene expansions were also observed in several other teleost fish species and were …

Tc-Ptp-Mediated Regulation Of P38 Mapk Pathway Promotes Uvb-Induced Apoptosis In Skin, Lindsey Susie Shim

Theses and Dissertations

T-cell protein tyrosine phosphatase (TC-PTP) is a non-receptor PTP that has been shown to have various roles in signaling pathways. Our previous studies have demonstrated that the TC-PTP deficiency in the epidermis can exacerbate hyperplastic response by inducing epidermal cell proliferation in response to tumor promoter TPA or UVB exposure. This implies that TC-PTP plays a tumor-suppressive role in the epidermis and provides protection against UVB radiation, which is well known to contribute to skin cancer development. The p38 mitogen-activated protein kinase (MAPK) is one of serine/threonine kinases that are activated through environmental stress including UVB. It has been demonstrated …

Effect Of An Anti-Hiv Polymer Based Nanoformulation On Immune Mediating Cells Within The Gut Associated Lymphoid Tissue And Its Behavioural Impacts, Blessed Ofori

Theses and Dissertations

This thesis investigates the therapeutic potential of nanoformulated antiretroviral therapy (NF cART) and its implications for HIV-associated chronic inflammation and neurobehavioral outcomes. In the first study, NF cART demonstrated enhanced efficacy in modulating inflammasome activation and reducing viral reservoirs in gut-associated lymphoid tissue (GALT) and the small intestine of HIV-infected humanized mice. Key markers of inflammation (NLRP3, Caspase-1, and IL-1β) and viral replication (LTR and p24 levels) were significantly suppressed with NF cART compared to conventional therapy, though residual LTR expression persisted in the small intestine. In the second study, nanoformulated Emtricitabine uniquely enhanced locomotor activity and reduced anxiety-like behaviors …

Investigation Of Reproductive Inhibition During Chronic Heat Stress In C. Elegans, Macy Makenna Nielander

Theses and Dissertations

Nearly all organisms experience heterogeneity in environmental conditions that can negatively impact fitness. A common adaptive strategy to reduce the impact of this stress is called facultative diapause, where development and reproduction are temporarily arrested to promote survival. Facultative diapause is characterized by three major criteria, the arrest must be (i) induced by an unfavorable environmental condition, (ii) reversible, and (iii) driven by genetically regulated mechanisms. The aim of this thesis was to determine whether chronic heat-induced inhibition of egg laying in Caenorhabditis elegans represents a novel form of facultative diapause. It was found that the inhibition of egg-laying is …

Dissecting The Interplay Of Protein Synthesis And Degradation Pathways In Cellular Adaptation To Stress, Brittany Friedson

Theses and Dissertations

Adaptation to stress requires cells to reprogram transcription, translation, and proteolytic pathways. Although much is known about the response of each program, it remains unclear how they coordinate following stress. My studies in S. cerevisiae identified the Cdk8 kinase module (CKM) of the Mediator complex as a new player in coordinating these processes. It is well established that the CKM consists of four highly conserved proteins (cyclin C, its cognate kinase Cdk8, and two structural proteins Med12 and Med13) and predominantly represses a subset of stress responsive genes in yeast. We demonstrated for the first time that the CKM also …

Developing Coexpression Systems To Introduce Hydroxyproline Into Protein Engineered Collagen Peptides Utilizing Hydroxylase From Acanthamoeba Polyphaga Mimivirus, Jennifer Soldatich

Theses and Dissertations

A major challenge of developing collagen mimetic peptides (CMPs) by bacterial expression is to include hydroxyproline for biomedical applications. Coexpression of Prolyl-4-hydroxylase from A.mimivirus with CMPs was investigated. Additionally, four expression designs were created for potential applications in mammalian cells to utilize their natural ability of proline-hydroxylation by post-translational modification.

Pm2.5 In The Respirable Air A Threat To Heart Health Investigating The Influence On Cardiovascular Performance And Unmasking The Potential Strategies For Cardio Protection, Bhavana Sivakumar

Theses and Dissertations

Air pollution, characterized by the presence of harmful substances in the atmosphere, represents a significant environmental and public health challenge impacting millions worldwide. The World Health Organization (WHO) reports that approximately 90% of the global population resides in regions where air pollution surpasses the recommended thresholds for healthy air quality. Within the realm of air pollution, the composition of particulate matter emerges as a pivotal consideration, particularly regarding PM2.5, comprising minute particles originating from diverse sources such as combustion processes, industrial operations, and natural phenomena like wildfires and dust storms. PM2.5 exhibit a range of chemical and physical attributes contingent …

Informatics-Based Discovery Of New Natural Products, Their Biosynthesis, And Their Biological Roles, Ethan Older

Theses and Dissertations

In this work, genome mining and biosynthetic knowledge are applied for the discovery of new natural products, their biosynthetic pathways, and their promising biological activities and roles through the development of two unique new informatics-based approaches.

In chapter one, an overview of microbial natural product biological functions and biosynthesis is provided. The use of bioinformatics leveraging microbial natural product biosynthetic knowledge is also introduced. This chapter highlights the importance and need for developing innovative approaches to accessing microbial natural products, a vastly untapped source of new molecules with promising biological activities.

In chapter two, a correlational network linking the gene-encoded …

Quantification Of Immunometabolite Derived Protein Modifications During Mitochondrial Metabolic Stress, Justin Hunter Cox

Theses and Dissertations

The immunometabolite itaconate accumulates during lipopolysaccharide (LPS) stimulation of macrophages and microglia. Itaconate non-enzymatically reacts with cysteine residues to generate 2,3-dicarboxypropylcysteine (2,3-DCP), referred to as protein dicarboxypropylation. The tricarboxylic acid (TCA) cycle metabolite fumarate non-enzymatically reacts with the amino acid cysteine to form S-(2-succino)-cysteine (2SC), resulting in irreversible protein succination. Since fumarate and itaconate levels dynamically change in activated immune cells, the levels of both 2SC and 2,3-DCP reflect the abundance of these metabolites and their capacity to modify protein structure and function. I generated esters of 2SC and 2,3-DCP from protein hydrolysates and used stable isotope dilution liquid chromatography-tandem …

Design Of An Artificial Transcription Factor For Inducing Potent Interferon-Γ Signals, Ashley King

Theses and Dissertations

Cytokines are small immune signaling proteins responsible for cellular communication and their levels are tightly regulated throughout immune recognition and defense. Interferon gamma is a key cytokine responsible for upregulation of antigen presentation and supports the differentiation of pro-inflammatory effector cells. Due to its role in regulation, excess interferon gamma is cleared from the body limiting the success of interferon gamma immunotherapies. This study reports the design and validation of an artificial transcription factor (ATF) that induces endogenous interferon gamma expression. The transcription factor contains zinc finger proteins to recognize the interferon gamma gene and an activation domain to promote …

Characterizing The Role Of The Escherchia Coli Monothiol Glutaredoxin, Grxd, As An Iron-Sulfur Cluster Carrier Protein, Claire Elizabeth Fisher

Theses and Dissertations

Iron-sulfur (Fe-S) clusters are important cofactors with a primary role of electron transport that are required by proteins involved in wide range of biological pathways such as respiration, photosynthesis, and DNA repair. In Escherichia coli (E. coli), the assembly and transfer of these clusters to their target apo-protein is a highly coordinated process encoded by the multiprotein systems iscRSUAhscBAfdx and sufABCDSE operons. These clusters are assembled on a scaffold protein, where they are then transferred to a carrier protein that will deliver the cluster to the target apo-protein. The Isc and Suf systems contain their own carrier proteins IscA …

Exploring The Role Of Rb1 In Cellular Differentiation Using Patient-Derived Stem Cells, Ambily Vincent

Theses and Dissertations

The Retinoblastoma (RB1) gene, predominantly recognized as a tumour suppressor, plays a crucial role in early growth and development. Mice lacking the retinoblastoma gene do not survive and exhibit defects in the differentiation of various tissues, underscoring its involvement in cellular differentiation. This suggests that the functions of the RB1 gene extend beyond its role as a tumour suppressor, encompassing cell cycle regulation, maintenance of genomic stability, cellular senescence, and cellular differentiation.

While the tumour suppressor function of the RB1 has been extensively studied, its role in cellular differentiation remains incompletely understood. Existing literature, primarily based on animal models, has …

Assessment Of Tripeptides Self-Assembly & Er Stress On Cell Viability And Exosome Secretion In Mda-Mb231, Azmat Parveen

Theses and Dissertations

This study delves into the effects of tripeptides KYpF and WYpK(NBD) on MDA-MB-231 cells, uncovering KYpF's ability to reduce cell proliferation and exosome secretion in peptide treated cells. Conversely, WYpK(NBD) demonstrated significant toxicity at 10 μM. These insights pave the way for optimizing peptide-based cellular treatments for exosome secretion purposes.

Documenting The Spatial And Temporal Dynamics Of Pair-Rule Genes In The Early Tribolium Embryo, Hasan Mahmud

Theses and Dissertations

Although Insects and vertebrates have different anatomical structures, they possess similar mechanisms for segmenting during embryogenesis. Clock and wavefront model exhibit a similar pattern of division as observed in Drosophila, like vertebrates, which emphasizes fundamental principles in the field of developmental biology. In contrast to Drosophila, the insect Tribolium castaneum exhibits posterior segmentation originating from the growth zone. The pair-rule genes’ expression is characterized by oscillatory patterns, which are controlled by a segmentation clock. Researchers found that the speed regulation model combines oscillatory and sequential gene activities to shape both elongating and non-elongating embryonic tissues. The adaptability of this model …

Interleukin 24 Induces Apoptosis Through Glycogen Synthase Kinase-3 Beta Inactivation In Prostate Cancer Cells, Sual J. Lopez, Moira Sauane

Theses and Dissertations

Interleukin 24 (IL-24) is a tumor-suppressing protein currently in clinical trials. IL-24 induces cancer-specific apoptosis by activating endoplasmic reticulum (ER) stress and mitochondria dysfunction. We have previously demonstrated that IL-24 leads to apoptosis in cancer cells by protein kinase A (PKA) activation in human breast cancer cells. To further understand the mechanism by which IL-24 induces apoptosis, I analyzed the role of glycogen synthase kinase-3 (GSK3), a highly conserved and normally active serine/threonine kinase in cancer cells and downstream target of PKA. The work reported here provides direct evidence that GSK3 was inhibited following IL-24 treatment in human prostate cancer …

Development And Characterization Of Β-Cyclodextrin Inclusion Complexes With Enzalutamide For Easing Administration And Efficacy Of The Drug In Crpc, Meghana Kolli

Theses and Dissertations

Prostate cancer (PC) is prevalent among men in the US, often leading to castration-resistant prostate cancer (CRPC) with therapy. This is marked by androgen receptor (AR) overexpression, resistance, mutations, and splice variants. Enzalutamide (ENZ), a nonsteroidal antiandrogen, surpasses first-gen AR inhibitors like Bicalutamide. It's the go-to drug for CRPC. We explore ENZ for self-assembly complex formation, crucial for CRPC treatment. Using the solvent evaporation method, we developed β-cyclodextrin and ENZ self-assembly complexes (β-CD-ENZ). β-CD acts as both agent and solubilizer, FDA-approved and safe. Efficacy was tested on PC cell lines (C4-2B) and (22Rv1), including 3D cultures for cytotoxicity and a …

Decisions, Decisions: Dna Damage Induced Modification Of P53 And The Effect Of Modifications On P53 Differential Binding To The Dna, Savannah Gabrielle Keating

Theses and Dissertations

p53 is the most frequently mutated tumor suppressor protein in cancer. It is highly regulated, most often via post-translational modifications (PTMs). Different pathways and target genes are differentially activated by p53 and there is much left to understand how this occurs. The regulation of p53 at the post-translational level is incredibly important to its DNA damage response, and, therefore, it is understood that PTMs are often involved in the differential activation of pathways by p53. This project aimed to identify some of the types and locations of post-translational modifications on p53 in response to different modes of DNA damage with …

Effect Of Plumbagin On Chemo-Resistant Metastatic Retinoblastoma, John J. Soto

Theses and Dissertations

Retinoblastoma, which is an ocular malignancy, usually results in poor prognoses in pediatric patients worldwide. Retinoblastoma in some events, can develop metastatic phenotypes which can lead to secondary tumor formation, furthering deleterious patient outcomes. It is of paramount importance to identify and research potent novel compounds that can be used to increase the likelihood of remission. Plumbagin (PLB) is a plant-derived, neuroprotective agent, which exhibits significant anticancer activities during in vitro study. PLB has been shown to have a high therapeutic efficacy against chemoresistant sublines as well as their normal counterparts. We attempted to show that the chemoresistant ABCC1 could …

Discovery Of Oligomeric States And Small Molecules That Affect Deacylase Activities Of Human Sirtuin-2 (Sirt2), Jie Yang

Theses and Dissertations

Human sirtuin isoform 2 (SIRT2) is an NAD+-dependent enzyme that removes various acyl modifications from lysine residues. SIRT2 is involved in many biological processes, such as transcription, DNA repair, cell proliferation, and metabolism. Due to its diverse functions, SIRT2 is implicated in various disease states, including cancers and neurodegenerative disorders. This dissertation aimed to understand the regulation of SIRT2’s deacylase activities and to discover small molecule modulators of SIRT2. In this work, we found that SIRT2 dimerized in solution, and the dimerization decreased the deacetylase activities without affecting the defatty-acylase activity of this protein. Dimerized SIRT2 dissociates into monomers upon …

Using Rapid Protein Degradation To Determine The Effect Of Runx1 Loss-Of- Function On Dna Damage Accumulation And Repair., Jackriel Pina Morales

Theses and Dissertations

Germline mutations in RUNX1 are associated with familial platelet disorder with a predisposition to myeloid malignancy (RUNX1-FPDMM), in which patients present with low platelet counts,excessive bleeding and bruising, and an increased risk of Acute Myeloid Leukemia (AML)/Myelodysplastic Syndrome (MDS) development throughout their lifetime. To understand how these loss-of-function mutations in RUNX1 drive predispose to malignancy, it is important to develop a detailed understanding of the molecular basis of RUNX1 function. While RUNX1 is a transcription factor, preliminary data from our group and work from others suggests that RUNX1 also interacts with proteins that are critical for DNA damage …