Biochemistry, Biophysics, and Structural Biology Commons^™

N-Allylation Of Secondary Amides With Unsymmetrical Propionates, Remi Graff

Honors Theses

The combination of trimethylsilyl trifluoromethanesulfonate (TMSOTf) and Et3N promotes the N-allylation of 4-methoxyacetanilide 2 with unsymmetrical propionates 1 (eq 1). The amide substrate is believed to undergo conversion to a nucleophilic silyl imidate in the presence of TMSOTf and the amine base, which concurrently promotes carbocation formation in the same flask through the loss of trimethylsilyl propionate, leading to N-allylation of the amide. A range of unsymmetrical propionates, accessed by varying the R and R’ substituents, react efficiently in this one-pot process with yields ranging from 44% to 99%, affording regioselective ratios of up to >20:1 through formation of regioisomers …

Development Of A Python Pipeline For The Inference Of Selection In Human Genomes And Applications To Adh1b*2, Aine Macdermott

Honors Theses

Human evolutionary genomics provides an avenue to understand how genetic variation has been impacted by the environmental pressures of the past, revealing how selective pressures impacted past populations and contributed to present-day biological diversity. In this thesis, I describe the construction of a Python-based computational pipeline designed to automate and integrate multiple analyses of positive selection across worldwide population datasets, including derived allele frequency calculation, haplotype-based tests of selection, RELATE-based genealogical inference, and CLUES2-based temporal modeling. By automating file preparation, format conversion, job submission, result aggregation, model comparison, and visualization, this pipeline streamlines the data interpretation process, and shifts focus …

Exploring Phosphoregulation Of Myo3a Using Quantitative Fluorescence Image Analysis In Cos7 Cells, Vu Hao Minh Nguyen Phan

Honors Theses

Myosin 3A (MYO3A) is an unconventional myosin involved in the formation and maintenance of hair-cell stereocilia of the sensory epithelia in the inner ear. The kinase domain has been implicated in phosphoregulation of MYO3A activity through intermolecular autophosphorylation. Previous studies using mass spectrometry identified two potential phosphorylation sites in the motor domain. To investigate the regulatory roles of these sites, we generated glutamic acid point mutations in our mchr-MYO3AΔK construct to mimic phosphorylation and assayed the constructs for their ability to tip-localize and influence filopodial density via transfection into COS7 cells. The phosphomimic constructs were less able to generate filopodia …

Biochemical Investigations Of F420-Dependent Glucose-6-Phosphate Dehydrogenase From Cryptosporangium Arvum, Sarah T. Al-Noubani

2026 Spring Honors Capstones Projects

F₄₂₀-dependent glucose-6-phosphate dehydrogenase (FGD) catalyzes the conversion of glucose-6-phosphate to 6-phosphogluconolactone using the deazaflavin, Cofactor F₄₂₀. FGD from Mycobacterium tuberculosis has been investigated due to its health relevance to prodrug activation. More recently, FGD from Cryptosporangium arvum (Cryar- FGD) has been investigated due to its broader substrate specificity toward multiple sugar phosphates. Kinetic and NMR studies have shown that Cryar-FGD has dual catalytic activity as a dehydrogenase and isomerase. Further investigations are being conducted on Cryar-FGD to characterize key residues involved in catalysis.

To investigate the catalytically active residues of Cryar-FGD, …

Systematics Of Disk-Winged Bats (Thyroptera), Courtney L. Klumpp

Capstone Showcase

This thesis investigates the evolutionary relationships among disk-winged bats in the genus Thyroptera, a small group of Neotropical bats known for the adhesive suction disks on their wings and feet that allow them to roost inside tightly rolled leaves. The distinctive morphology and behavior of these bats have not been extensively documented, and their phylogenetic relationships remain only partially resolved. The goal of this research is to better understand how these species are related to one another and to determine whether the five recognized species of Thyroptera form a single monophyletic group. To address this question, morphological and genetic …

Seeing Through The Fuzz: Understanding Conformational Diversity In The Fuzzy Coat Of Amyloids Across Fibril Polymorphs Using Solid-State Nmr, Carah K. Allen

Scripps Senior Theses

The aggregation of α-synuclein (α-syn) into amyloid fibrils is a central pathological hallmark of Parkinson’s disease and other related synucleinopathies. Although monomeric α-syn is intrinsically disordered, the protein assembles into multiple fibril polymorphs that differ in their β-sheet core structures. Surrounding each polymorph is the “fuzzy coat,” which is a highly dynamic, intrinsically disordered region (IDR) that is not resolved by high-resolution structural techniques, such as Cryo-EM, but is thought to influence fibril interactions, propagation, and toxicity. Understanding whether different polymorphs maintain distinct fuzzy-coat conformational ensembles is therefore essential for connecting amyloid structure to biological function. To address this question, …

How Semiflightless Birds Adapt: A Survey From Barú, Costa Rica, Eli P. Laloudakis

Scripps Senior Theses

When we think of birds, we think of flight. Because of this, scientific research often focuses on how birds evolved flight and how their wings function during flight. However, not all birds can fly or fly well, as wings wax and wane on developmental, seasonal, and evolutionary time scales (Dial and Heers 2021). Consequently, rudimentary wings can be found in developing birds, secondarily flightless or semiflightless birds, birds that undergo simultaneous molts, and extinct theropods with “protowings” (structures that preceded fully functional wings). Secondarily flightless and semiflightless birds often evolve where there is no/low predation, abundant year-round food supply, and …

Overview And Methods Of Molecular Paleontology: A Review, Shelby Touchstone

Williams Honors College, Honors Research Projects

Molecular paleontology—the study of ancient biomolecules preserved in fossils and sediments, including ancient DNA, lipids, and proteins—has motivated researchers since the 1950s, inspiring both scientific inquiry and broader cultural imagination. Think Jurassic Park. This honors thesis presents a comprehensive review of the history, current state, and methods of the field of molecular paleontology. Drawing on peer-reviewed literature and scholarly resources accessed through databases such as EBSCO and Bierce library, this review examines key techniques such as Next Generation Sequencing and amino acid racemization and will go over limitations and controversies in the field. It will also explore potential new …

Structural Determination And Regulatory Mechanisms Of Rice Lipoxygenase-1, Lillie Ballanco, Nathaniel Gilbert, Adam Offenbacher

Distinguished Undergraduate Researcher Program

Lipoxygenases (LOXs) are non-heme iron containing enzymes that catalyze the oxygenation of polyunsaturated fatty acids (PUFAs) such as linoleic acid (LA). These lipoxygenase enzymes are found in both plants and animals. In humans, LOXs have been linked to inflammatory diseases such as asthma, atheroscelerosis, and allergic rhinitis. Plant LOXs have been primarily linked to the growth, development, and stress response of the plant. This paper focuses on Oryza sativa lipoxygenase-1 (OsLOX1), an enzyme found in rice (Oryza sativa). OsLOX1 has been associated with enhanced drought tolerance in rice and may play a role in regulating growth under water-limited conditions. Understanding …

An Investigation Into The Potential Role Of Thioester Bonds In Prebiotic Atp Synthesis, Adam J. Ruf

Williams Honors College, Honors Research Projects

In this project I will attempt to show a potential prebiotic pathway for the synthesis of ATP. This pathway involves the use of thioester bond containing molecules, as derivatives of these bonds have been shown to exhibit enzyme-like catalytic properties. Nuclear magnetic resonance spectroscopy will be utilized to characterize each step of the pathway to ensure it is successful.

C. Difficile Detection Method For First Responder Glove Application, Alli N. Senedak

Williams Honors College, Honors Research Projects

Clostridioides Difficile (C. diff) is a Anaerobic Gram-positive bacillus that is capable of spore formation, making it difficult to control its spread and duration in clinical environments. This phenomenon can provide danger to first responders, healthcare workers, and patients. The goal of this project is to create a biosensor capable of detecting C. diff in a clinical setting that can be applied to a glove apparatus. The project will involve the use of C. diff aptamers activated on the surface of an electrode. Once surface activation has been verified via surface analysis, the electrodes will be exposed to C. diff …

Multicomponent Deoxyribozyme Probes For Targeted Detection Of Cancer Associated Gene Insertions, Samuel R. Veleke

Honors Undergraduate Theses

Insertions and deletions (indels) in DNA sequences of genes are linked with cancers such as glioblastoma (GB). Thus, indels may be used as potential cancer biomarkers. In the NANOG gene, its pseudogene NANOGP8 has a 22 base pair insertion sequence for GB patients. In this work, the NANOG gene and the insertion sequence were detected in one sample using a dual-fluorescent assay containing two multicomponent split deoxyribozyme (Dz) probe complexes. One complex used TXR fluorescence to detect the gene regardless of the insertion’s presence. The other complex used fluorescein (FAM) fluorophore to detect the insertion. The probes were able to …

Bio-Orthogonal Chemistry-Based Strategy To Turn-On And Turn-Off Crispr-Cas9 Gene Editing In Solution And In Live Cells, Bhoomika Pandit

Electronic Theses & Dissertations (2024 - present)

The CRISPR–Cas9 system is a widely popular tool for genome engineering. There is a strong interest in developing tools for temporal control of CRISPR-Cas9 activity to address some of the challenges and to broaden the scope of potential applications. In this thesis I describe two biorthogonal based approaches to either Turn-ON and Turn-OFF CRISPR-Cas9 gene editing.

In first project work I describe a bio-orthogonal chemistry-based approach to Turn-ON Cas9 nuclease activity with temporal precision. We report a TCO-acylimidazole reagent that acylates 2′-OH groups of RNA. Poly-acylation (“cloaking”) of RNA was optimized in vitro using a model 18-nt oligonucleotide, as well …

Disrupted Circadian Rhythms Affect Hallmark Pathologies In Alzheimer’S Disease-Related Mouse Models, Valeria Buzinova

Theses and Dissertations--Molecular and Cellular Biochemistry

Alzheimer’s Disease (AD) is a complex neurodegenerative disease with two hallmark pathologies: extracellular amyloid-b (Ab) and intracellular neurofibrillary tangles (NFTs). Ab is proteolytically processed from amyloid precursor protein (APP) by b-secretase and g-secretase as a monomeric peptide prone to aggregation. Eventually, the aggregate-prone monomers will form dense plaques that are difficult to break down and remove. These plaques begin to deposit into the cortex decades prior to the formation of NFTs and the onset of cognitive decline. NFTs are comprised of hyper-phosphorylated tau. Tau is a protein that serves to promote and stabilize the formation of microtubules. The formation of …

Characterizing The Human Metapneumovirus Matrix Protein And Its Role In Infection, Chase J. Heim

Theses and Dissertations--Molecular and Cellular Biochemistry

Human metapneumovirus (HMPV) causes severe respiratory tract infections in all cohorts, but especially in vulnerable groups such as children, older adults, and the immunocompromised. The matrix (M) protein of HMPV, like the M proteins of other members of the Mononegavirales order, is involved in virus assembly and budding. However, other functions of the HMPV M protein have yet to be elucidated. To investigate these various functions of the M protein, we used a peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) antisense agent designed to block translation of the M gene to reduce M expression during infection. Treatment with varying concentrations of an …

Absolute Quantification And Identification Of Rna From Rna-Lipid Nanoparticles Using High Resolution Mass Spectrometry, Jason C. Funderburk

Theses and Dissertations

RNA therapeutics are a rising drug category with potential use for a range of conditions encompassing infectious diseases to therapies for cancer, diseases, and genetic disorders. RNA-lipid nanoparticles (RNA-LNPs) are the prominent delivery method for these therapeutics approved products include mRNA vaccines and polyneuropathy treatments. The emergency use authorizations and orphan drug status of current RNA-LNP drugs has allowed approval without finalization of the regulatory analytical procedures for quality monitoring. The objective of the study was to develop an LC-MS assay to simultaneously measure identity and concentration of two therapeutically relevant intact RNA constructs extracted from RNA-LNPs to enhance quality …

Interaction Of The Dap-10 Fluorescent Light-Up Aptamer With An Environment-Sensitive Dye, Crystal Violet, Marina Tsukanova

Honors Undergraduate Theses

Crystal violet (CV) is an environment-sensitive dye whose fluorescence is strongly influenced by microenvironment polarity and viscosity, increasing markedly when rotational and vibrational motions are restricted. While the fluorescence enhancement of CV by fluorescent light-up aptamers (FLAPs) has been noted in literature, the specific binding interactions between CV and aptamers in the DAP-10 family have remained under-investigated.  The objective of this work is to elucidate the binding site interactions between CV and the DAP-10-42 FLAP. It is hypothesized that positions 34 and 35 of the aptamer’s sequence reside in a proposed quadruplex-duplex junction that form the dye-binding pocket of DAP-10 …

Exploring Anticholinergic Neurotoxicity: Diphenhydramine's Impact On Neuro-2a Cell Morphology, Expression, And Viability, Jordan B. Mcintyre

Honors Undergraduate Theses

Anticholinergic medications, such as diphenhydramine (commonly known as Benadryl), are increasingly recognized for their association with cognitive impairment and neurodegenerative disease. These medications, which inhibit the action of acetylcholine, a neurotransmitter essential for the formation and utilization of the central and peripheral nervous system, have been implicated in conditions such as dementia and may impact fundamental cellular processes like neurogenesis and cell proliferation. This study investigates the effects of diphenhydramine on neurogenesis when introduced to Neuro-2a cells during differentiation. For all studies cells were plated on coverslips or in 6-well plates with differentiation media containing 0 ng/mL, 50 ng/mL, …

Development Of A Split Deoxyribozyme-Based Assay For Detecting Hepatitis B Virus, Dalila Zovko

Honors Undergraduate Theses

Rapid, accessible nucleic acid-based detection methods are needed for hepatitis B virus (HBV). A split deoxyribozyme assay was developed for sequence-specific fluorescence detection. The assay was evaluated using synthetic target and plasmid-derived material. The assay showed target-dependent fluorescence and low-concentration linear range with a LOD of 1.9 nM. The assay supports detection and semi-quantification of HBV genetic material as shown using plasmid-derived transcript samples from HBV genotypes B, C, and D. The selected target corresponded to nt 363–406 of the HBV genome and was chosen based on sequence alignment showing conservation across most genotypes. Genotype-specific detection remains a future application …

Dna Half-Adder Based Calculator Driven By 4j Molecular Switches, Andrea Gissele Canizalez Covaleda

Honors Undergraduate Theses

DNA nanostructures have computing functions due to the predictable base pairing of nucleic acids (Watson–Crick). This property allows the programming of DNA circuits that integrate DNA logic gates capable of recognizing oligonucleotides. This molecular device offers the advantages of biocompatibility and potential applications in personalized medicine for diagnosis and therapies.

Our lab has developed DNA logic gates formed by the connection of YES strands via DNA four-way junctions (4J), which can function as molecular switches to develop a DNA-based calculator. Our DNA half-adder calculator nanostructure is designed to recognize up to three combinations of DNA inputs using three DNA strands …