Biochemistry, Biophysics, and Structural Biology Commons^™

Investigating The Ability Of Presequences To Influence Mitochondrial Protein Import, Youmian Yan

WUSM Theses and Dissertations – All Programs

Mitochondria are essential organelles housing over 1000 proteins, 99% of which are nuclear-encoded and synthesized in the cytosol. Most mitochondrial proteins are targeted by a positively charged sequence called the presequence, which lacks a common sequence motif and is poorly conserved across evolution. Accumulating evidence suggests that presequences encode meaningful information that influences mitochondrial protein uptake. For instance, multiple stress response models propose that stress sensing proteins contain “weak” presequences which allows them to monitor mitochondrial health. However, there is a lack of systematic examination about how presequences influence import kinetics and what constitutes presequence strength. These gaps in knowledge …

The Β2Αb Loop Determines Nad(P) Cofactor Specificity And Kinetics In Trypanosomal D-3-Hydroxybutyrate Dehydrogenases, Hideharu Hashimoto, Ian H. Mawn, William Escobar-Arrillaga, Linh Nguyen, Laura A. Madigan, Gina Antuono, Tatiana Rossy, Jorna Sojati, Anna Mienko, Erik W. Debler, Jennifer B. Palenchar

Department of Biochemistry and Molecular Biology Faculty Papers

Bacterial d-3-hydroxybutyrate dehydrogenases (HBDHs) catalyze the conversion between d-3-hydroxybutyrate and acetoacetate with NAD as the cofactor but not with NAD 2'-phosphate (NADP). However, HBDHs of the early-branched eukaryotic genus Trypanosoma utilize both NAD and NADP (T. brucei) or exclusively NADP (T. cruzi). Here we reveal that NADP specificity of T. cruzi HBDH arises from stabilization of the flexible β2αB loop by the 2'-phosphate interaction. Stabilization of this loop by a nearby C64Y mutation enables T. cruzi HBDH to use NAD in addition to NADP; thus, the Cys/Tyr residue is critical for determining cofactor specificity in trypanosomal HBDHs, suggesting that most …

Evaluation Of Polycyclic Aromatic Hydrocarbon Profiles In Fermented Artemisia Annua Leaf Extract, Mabitine Daniel Malo, Ohemu Adaofoyi Isaiah, Igaznda Terhemba James, Obamuwe Rebecca Joy, Ugber Erdoo Charity, Itodo Monday Christopher

Journal of Bioresource Management

This study evaluated the presence, profile and concentration of polycyclic aromatic hydrocarbons (PAHs) in the fermented leaf extract of Artemisia annua, a medicinal plant widely used for its antimalarial and skin-benefiting properties. Given the plant’s increased use in Nigerian traditional medicine, this research examines how fermentation a process known to enhance bioactive compound availability may influence PAH levels, which are of concern due to their carcinogenic, mutagenic, and teratogenic effects. The A. annua leaves were fermented using Saccharomyces cerevisiae cultures and subsequently analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) to detect and quantify PAHs. The analysis identified 24 distinct PAH …

Evaluation Of The Dhcr7/Nadsyn1 Rs4944946 Variant And Breast Cancer Susceptibility: A Case-Control Study In An Indonesian Cohort, Dian Tri Wahyudi, Rafika Indah Paramita, Sonar Soni Panigoro

Indonesian Journal of Medical Chemistry and Bioinformatics

Background: The rs4944946 variant, located in the 5’UTR of the DHCR7 gene and upstream of NADSYN1, is biologically implicated in vitamin D and cholesterol metabolism pathways closely linked to cancer pathogenesis. This study aimed to experimentally validate the association between the rs4944946 polymorphism and breast cancer susceptibility in an independent cohort of Indonesian women. Method: A case-control study was conducted involving 36 breast cancer patients and 39 healthy adult females. Genotyping of the rs4944946 variant was performed using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP). The genetic association was evaluated across codominant, dominant, and recessive models. Luminal B was the …

Development Of An Indirect Elisa Kit For Measurement Of Circulating Thyroid Peroxidase Levels In Patients With Hypothyroidism, Aulanni’Am Aulanni’Am, Devi Ramadani, Almas Dwi Khairana, Saidun Fiddaroini, Muhammad Fikri Nur, Andreas Budi Wijaya, Rulli Rosandi, Dyah Kinasih Wuragil, Wibi Riawan

Makara Journal of Science

Hypothyroidism, characterized by insufficient thyroid hormone production, affects approximately 5% of the population, especially women, thus increasing pregnancy-related risks. Conventional screening mainly measures thyroid-stimulating hormone (TSH) and free thyroxine (FT4). Detecting the presence of anti-thyroid peroxidase (anti-TPO) antibodies indi-cates autoimmune thyroid disease. This study developed an indirect enzyme-linked immunosorbent assay (ELISA) kit for detecting circulating TPO using polyclonal antibodies generated in rabbits. TPO was isolated from positive sera of pa-tients with hypothyroidism, purified by electroelution, and confirmed as a 52-kDa protein by Western blot. Rabbits immunized with this protein produced polyclonal antibodies with the highest titer (OD = 0.334) at …

Bambara Groundnut (Vigna Subterranea L.) Supplementation Ameliorates The Histological Structure Of Skeletal Muscle In Protein-Deficient Mice, Raquellynda Fadilla Syafanisa, Rimonta Febby Gunanegara, Sunarti Sunarti, Siti Aeniah, Ardaning Nuriliani

Makara Journal of Science

Protein deficiency may cause histological alterations and pathological effects in skeletal muscle, leading to reduced mus-cle size and functional degeneration. Bambara groundnuts contain essential amino acids that have the potential to meet the body’s protein requirements. Therefore, this study aimed to investigate the effect of bambara groundnut supplemen-tation on skeletal muscle in protein-deficient mice. Twenty-five female mice were divided into five groups (control, pro-tein deficiency, and protein deficiency with 100, 200, and 300 g of bambara groundnut supplementation/kg of feed). The treatment was administered for 2 months. At the end of the experiment, the left gastrocnemius muscles of the mice …

Ddrr Acts As A Dna Damage-Induced Cell Division Inhibitor In Acinetobacter Baumannii, Christian P. Lowe, Deborah Cook, Lily Hoard, Mckinnley Martin, Janelle M. Hare

Acinetobacter baumannii DNA Damage Database

An article titled, "DdrR acts as a DNA damage-induced cell division inhibitor in Acinetobacter baumannii" published in the Journal of Bacteriology on September 9, 2026.

Cultivating Collaboration In Biomolviz: Key Attributes For Building A Stem Educator Community Of Practice, Pamela S. Mertz, Charmita Burch, Roderico Acevedo, Josh T. Beckham, Didem Vardar-Ulu, Kristin M. Fox, Lauren A. Genova, Rachel M. Mitton-Fry, Swati Agrawal, Daniel R. Dries, Kristen Procko

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

BioMolViz is an educational community of practice (CoP) focused on creating resources to improve biomolecular visual literacy instruction and assessment. The community of life science instructors developed in three phases, beginning with a small Core Team that emerged from a conference session and established the domain of interest for the CoP. In-person workshops, supported by short-term funding, drove the initial expansion of the community and the development of shared activities in Phase 2. This CoP was sustained through intentional choices about how members interact, collaborate, and share work. As the community matured in Phase 3, it was supported by longer-term …

Analysis Of Diterpenoid Alkaloids From American Aconitum Using Lc-Ms Metabolomic Methods, Yi Zhao

Dissertations, Theses, and Capstone Projects

Analysis of Diterpenoid Alkaloids from American Aconitum Using LC-MS Metabolomic Methods

by

Yi Zhao

Advisor: Edward J. Kennelly

Aconitum plants have been used as a traditional medicine in Asian countries for millennia to treat pain, rheumatism, syncope, fever, and other conditions. These therapeutic effects are mainly due to the bioactive diterpenoid alkaloids accumulated in the roots. Comparing with the large records and studies of Asian Aconitum and their medicinal use and the broad bioactive compounds, American Aconitum has been rarely investigated, not to mention how American Aconitum has been used medicinally. Based on previous phylogenetic and chemotaxonomic studies of Aconitum …

Waste Streams From Next Generation Molten Salt Reactors: The Challenge Of Creating An Insoluble Ceramic For Chloride Salt Waste, Alevtina A. Maksimova, Jake W. Amoroso, Matthew Page, Gregory Morrison, Hans Conrad Zur Loye

Faculty Publications

Molten salt reactors (MSRs) design development is of interest to many research groups and companies. Waste management for MSRs is one of the most important issues that needs to be resolved due to its impact on environmental safety, primarily via the dissolution of radionuclides in water. Furthermore, the waste treatment strategy will have a significant influence on the operating cost of MSRs, which is why a reliable process for the immobilization, transportation, storage and disposal of MSR waste must be addressed. This work presents a new approach for the immobilization of chloride salts from molten salt reactors in stable oxyhalide …

Sec-Saxs And Sans Analyses Of The Talin•Vinculin Complex, Gilbert Reyes

Dissertations, Theses, and Capstone Projects

Focal adhesions (FA) are integrin-based cell-matrix adhesion sites composed of adaptor proteins that facilitate the generation of traction forces important for biological functions such as cell migration, embryo development, wound healing, tissue organization and stability, and immune responses. Mature FA are observed when both autoinhibited multi-domain scaffolding protein talin and membrane-cytoskeletal protein vinculin are activated and interact thus promoting the assembly and function of FA. While research has resolved truncated sections of autoinhibited talin and recruitment of vinculin due to mechanotransduction that exposes cryptic vinculin binding sites (VBS), little is known about the global modifications of full length talin in …

Riluzole As A Dual-Targeted Radiosensitizer For Osteosarcoma: Targeting Tumor Cells And Angiogenic Vasculature To Enhance Single High-Dose Radiotherapy Efficacy, Pooja P. Rao

Dissertations, Theses, and Capstone Projects

Osteosarcoma (OS), the most common primary bone malignancy of children and young adults, is intrinsically radioresistant, and outcomes for patients with unresectable or metastatic disease have not improved in over four decades. Single-dose radiation therapy (SDRT) is a potent modality against radioresistant, highly vascularized sarcomas, but its application in OS is constrained by dose-dependent normal tissue toxicity. Radiosensitizers achieving tumor control at lower doses therefore offer a direct route to widening the therapeutic window. Critically, OS radioresistance is not tumor-cell-autonomous. It emerges from two coordinated compartments: aberrant DNA repair and antioxidant capacity intrinsic to the tumor cell, and a radioprotective, …

Characterization Of Mettl3/14-Mediated M6a Modification In Human Transcriptome Using Nanopore Direct Rna Sequencing, Emily Kurtyan, Andrew J. Stein, Kelly J. Abdalla, Zhangerjiao Yuan, Miten Jain, Fadia Ibrahim

Department of Biochemistry and Molecular Biology Faculty Papers

Post-transcriptional RNA modifications modulate diverse aspects of RNA metabolism. N6-methyladenosine (m6A), one of the most abundant internal RNA modifications, is deposited by the core methyltransferase complex, METTL3 and METTL14. Oxford Nanopore Technologies (ONT) platform permits direct, single RNA molecule sequencing while preserving native modifications. However, without rigorous benchmarking, the accuracy and reproducibility of modification detection remain uncertain. Here, we leveraged ONT to comprehensively profile bona fide m6A modifications in cellular RNAs at single-nucleotide resolution by integrating two direct RNA sequencing chemistries (RNA002 and RNA004) with the m6Anet and Dorado modification-detection models. We independently depleted METTL3 and METTL14 in human cells …

The Role Of External Factors In Shaping Carbohydrate Conformational Landscapes, Murat Yaman

Dissertations, Theses, and Capstone Projects

Carbohydrates (glycans) are among the most structurally diverse biomolecules and participate in a wide range of biological processes, including molecular recognition, cell adhesion, immune response, host-pathogen interactions, and cellular signaling. Their biological functions are governed by a complex interplay between sequence, stereochemistry, conformational flexibility, and environmental factors. However, the structural heterogeneity and dynamic nature of glycans present significant challenges for both experimental characterization and theoretical modeling. In particular, the relationships between glycan structure, conformational dynamics, and the resulting potential energy surfaces remain incomplete.

This dissertation investigates how external factors modulate the potential energy surfaces of carbohydrates through a combination of …

Evaluation Of Mdm2 Mrna Expression According To Clinicopathological Characteristics In Oral Squamous Cell Carcinoma (Oscc) Patients, Tirta Wardana

Chulalongkorn Medical Journal

Background: The murine double minute 2 (MDM2) gene encodes an E3 ubiquitin ligase that targets p53 for degradation, thereby influencing the pathogenesis of oral squamous cell carcinoma (OSCC).

Objective. The present study investigates MDM2 mRNA expression and its association with clinical and pathological characteristics in OSCC patients.

Methods: We conducted an observational-analytical study using a cross-sectional design. The study involved 39 OSCC patients. RNA extraction, cDNA synthesis, and quantitative PCR (qPCR) were performed. We quantified relative MDM2 mRNA expression using the Livak method (2-ΔΔCT) and GenEX MultiD software version 6.0. Statistical significance was assessed using independent t-tests.

Results: Although no …

Development Of Ultrafast Protein Digestion And Standards Free Quantitation Methods For Mass Spectrometric Analysis, Praneeth Ivan Joel Fnu

Dissertations

A wide variety of biologically important molecules, such as enzymes, antibodies, hormones, transporters and receptors are proteins by composition and they play key roles in biological functions such as cellular regulation, communication, metabolism and physiological function. Protein dysfunctions and abnormalities are associated with numerous diseases, making proteins critical targets for understanding disease mechanisms and developing therapeutic interventions. Protein-based therapeutics such as monoclonal antibodies, hormones and vaccines gained popularity due to their high specificity, efficacy, and ability to treat complex diseases that are often difficult to address with small-molecule drugs.

Given the important role of proteins in a variety of biological …

Modeling Kinetics Of 5-Aminolevulinic Acid-Induced Protoporphyrin Ix Accumulation For Optimal Photodynamic Therapy, Priscilla Melebor

Graduate Masters Theses

Photodynamic therapy (PDT) is a minimally invasive treatment that uses light-activated photosensitizers to selectively destroy diseased tissue through the generation of reactive oxygen species. However, conventional in vitro models often fail to accurately reproduce the pharmacokinetics of photosensitizer accumulation and clearance observed in vivo. This study investigated protoporphyrin IX (PpIX) accumulation following 5-aminolevulinic acid (ALA) exposure under physiologically relevant conditions using two-dimensional (2D) monolayers and three-dimensional (3D) spheroids. By comparing continuous ALA exposure with transient exposure followed by ALA washout, this work provides a more biologically relevant framework for studying PpIX kinetics and optimizing photodynamic therapy.

Synthesis, Utilization And Biological Evaluation Of Β-Hairpin Peptides As Inhibitors Of Immune Checkpoints, Sarah Naylon

Electronic Theses and Dissertations

Peptide therapeutics have emerged as promising alternatives to monoclonal antibodies because of their smaller size, improved tissue penetration, and synthetic versatility. Yet their broader application is often limited by poor structural stability, rapid clearance, and reduced target residence times. This dissertation describes the design, synthesis, structural characterization, and biological evaluation of β-hairpin peptides as inhibitors of immune checkpoints and as peptide vaccine scaffolds while developing strategies to overcome these limitations. At first a series of antibody-derived β-hairpin peptides (mimics of complementary determining region heavy-chain 3: CDR-H3s) as covalent inhibitors of the PD-1/PD-L1 immune checkpoint were investigated. Electrophilic warheads were incorporated …

Instruction Guide, Mark C. Hall

Analytical Biochemistry: A Practical Handbook for Undergraduates

The instruction guide provides useful information for setting up and running the lab activities each week and assessing student preparation, performance, and comprehension.

The Defining Role Of Ago4 And Its Catalytic Activity During De Novo Rna-Directed Dna Methylation, Tianyi Li

Arts & Sciences Graduate Student Theses and Dissertations

Plant transposable elements and transgenes are continuously targeted for transcriptional repression via the pathway of RNA-directed DNA methylation (RdDM). Although perpetual RdDM of the endogenous genomic loci has been well studied in the field, much less is known about the de novo RdDM pathway, the pioneering round of DNA methylation responsible for triggering and initiating silencing of new transgenes. Despite being two distinct pathways, both perpetual and de novo mechanisms of RdDM require the production of small interfering RNAs (siRNAs) and the key proteins of the ARGONAUTE4 (AGO4) clade. However, the order and likely mechanism of recruitment of AGO4 to …