Physical Sciences and Mathematics Commons^™

The Α-Crystallin Chaperones Undergo A Quasi-Ordered Co-Aggregation Process In Response To Saturating Client Interaction, Kirsten Lampi, Adam P. Miller, Susan E. O'Neill, Steve L. Reichow

Chemistry Faculty Publications and Presentations

Small heat shock proteins (sHSPs) are ATP-independent chaperones vital to cellular proteostasis, preventing protein aggregation events linked to various human diseases including cataract. The α-crystallins, αA-crystallin (αAc) and αB-crystallin (αBc), represent archetypal sHSPs that exhibit complex polydispersed oligomeric assemblies and rapid subunit exchange dynamics. Yet, our understanding of how this plasticity contributes to chaperone function remains poorly understood. This study investigates structural changes in αAc and αBc during client sequestration under varying degree of chaperone saturation. Using biochemical and biophysical analyses combined with single-particle electron microscopy (EM), we examined αAc and αBc in their apo-states and at various stages of …

From Grain To Malt: Tracking Changes Of Ultra-Low-Gluten Barley Storage Proteins After Malting, Mahya Bahmani, Angéla Juhász, Utpal Bos, Mitchell G. Nye-Wood, Malcolm Blundell, Crispin A. Howitt, Michelle L. Colgrave

Research outputs 2022 to 2026

Barley (Hordeum vulgare L.) is a major cereal crop produced globally. Hordeins, the major storage proteins in barley, can trigger immune responses leading to celiac disease or symptoms associated with food allergy. Here, proteomics approaches were employed to investigate the proteome level changes of grain and malt from the malting barley cultivar, Sloop, and single-, double- and triple hordein-reduced lines. The triple hordein-reduced line is an ultra-low gluten barley cultivar, Kebari®. Using discovery proteomics, 2,688 and 3,034 proteins in the barley and malt samples were detected respectively. Through the application of targeted proteomics, a significant reduction in the quantity …

Using Quantitative And Kinetic Proteomics To Explore Proteostasis, Nathan Raul Zuniga Pina

Theses and Dissertations

Every cell consists of carefully orchestrated biomolecules such as lipids, carbohydrates, and proteins. To maintain internal stability (homeostasis), cells maintain the right amount of these molecules at the right time and at the right place. This process is especially true for proteins since they are the foundation functional units within the cell. Proteins form structures and perform chemistry that bestows cells overarching functional roles. Cells maintain protein homeostasis (proteostasis) by modulating synthesis, folding, and degradation processes (turnover) to maintain the abundance levels for all proteins. This is the foundational kinetic model of proteostasis that is covered in this work, and …

Monitoring Brain Region-Specific Control Of Protein Turnover And Concentration Using Proteomics, Rebecca Suzanne Burlett

Theses and Dissertations

Regulation of metabolism is vital to health and lies at the core of many different diseases. The breakdown of metabolisms' regulation within the brain can lead to neurological disease like Alzheimer's Disease (AD). AD is known to affect brain regions responsible for memory and memory processing like the hippocampus and entorhinal cortex. The regulation of these regions' protein quality, synthesis, and degradation deviate from 'normal' or 'healthy' levels when AD is happening. It is known there is a breakdown of regulation in those regions; however, little is known about the specifics of regulation in healthy brains regions or how it …

Soft Contact Lens Tear Film Proteomics For Clinical Diagnostic Biomarker Discovery, Robert Kimball Roden

Theses and Dissertations

Human tear film is an accessible biospecimen rich with useful biological information. As tear protein biomarkers for both ocular and systemic diseases have been identified in tears, efforts are being made to create point-of-care tests useful for clinical diagnoses. However, there are significant obstacles to tear diagnostics, including inadequate sampling methods and biomarkers with insufficient sensitivity and specificity. Furthermore, tear film diagnostics are particularly challenging for dry eye disease (DED) patients, where low tear volume makes sampling more difficult and the multifactorial nature of DED makes identifying the exact pathological subtype very complex. As soft contact lenses (SCLs) are designed …

Linking Decomposition Reactions In Arctic Soils To Microbial Enzyme Production, Jane Karen Martinez

Open Access Theses & Dissertations

Microbial processes such as soil enzyme production are a major driver of decomposition and a current topic of interest in arctic soils due to the effects of climate warming. Despite the advances in understanding soil enzymes, there are still knowledge gaps regarding the role of enzymes in decomposition. In this dissertation, I addressed three of those gaps in the following chapters: (Ch.2) to explore the location of enzymes within the soil matrix, (Ch.3) to identify peptides matched to soil enzymes produced by microorganisms for organic matter decomposition, and (Ch.4) understand the longevity of enzymes in the soil after microbial production. …

Analysis Of Post-Translational Modifications (Ptm) Crosstalk, Amit Das

Theses and Dissertations

Mass spectrometry-based proteomics is a powerful tool for identifying post-translational modifications (PTMs) across the proteome. O-GlcNAcylation and phosphorylation are two PTMs that play crucial roles in regulating cellular processes, including cardiac contractile function. Dysregulation of these PTMs has been implicated in the development and progression of diabetic cardiomyopathy. In this study, we aimed to investigate the interplay between O-GlcNAcylation and phosphorylation in healthy and type 2 diabetic hearts, with a specific focus on the functional relationships between these PTMs and their potential therapeutic implications.

Utilizing mass spectrometry data, we identified and quantified specific PTMs on myofilament proteins, uncovering 1354 O-GlcNAcylated …

Quantitative Analysis Of The Proteomic Selectivity Of Acidic Reductive Alkylation Of Peptides, Connor Jewell

Honors Scholar Theses

Proteome quantification is a complicated process which takes on many methods, usually involving peptide derivatization and complex mass spectral analysis. Scores of techniques have been utilized to address issues of incomplete quantification. The implementation of IPTL labeling, or isobaric peptide terminal labeling, is a recent addition to the literature, which exploits the supposedly selective labeling of N-termini of tryptic peptides from the C-termini of lysine residues due to differences in basicity. This method has been shown to improve quantitation by reducing spectral complexity and provide more accurate and reproducible results on peptide abundance. Investigations into the selectivity of terminal derivatization …

Prediction Of Rapid Early Progression And Survival Risk With Pre-Radiation Mri In Who Grade 4 Glioma Patients, Walia Farzana, Mustafa M. Basree, Norou Diawara, Zeina Shboul, Sagel Dubey, Marie M. Lockheart, Mohamed Hamza, Joshua D. Palmer, Khan Iftekharuddin

Electrical & Computer Engineering Faculty Publications

Rapid early progression (REP) has been defined as increased nodular enhancement at the border of the resection cavity, the appearance of new lesions outside the resection cavity, or increased enhancement of the residual disease after surgery and before radiation. Patients with REP have worse survival compared to patients without REP (non-REP). Therefore, a reliable method for differentiating REP from non-REP is hypothesized to assist in personlized treatment planning. A potential approach is to use the radiomics and fractal texture features extracted from brain tumors to characterize morphological and physiological properties. We propose a random sampling-based ensemble classification model. The proposed …

Mass Spectrometry-Based Proteomic Analysis To Characterise Barley Breeding Lines, Mahya Bahmani

Theses: Doctorates and Masters

Barley is a key ingredient in the malting and brewing industry, and it is the fourth most important crop being cultivated worldwide. The protein content of the barley grain is one of the main components determining the quality and nutritive value of the food and beverages prepared from barley. Mass spectrometry-based proteomic analysis is a valuable tool that can guide and inform plant breeding strategies and crop improvement programs. Understanding the proteome changes in barley grain under different growing locations, the impact of different environmental conditions and its relationship with malting characteristics have the potential to inform breeding programs to …

Utilizing Proteomics To Identify And Optimize Microalgae Strains For High-Quality Dietary Protein: A Review, Sara Hamzelou, Damien Belobrajdic, James A. Broadbent, Angéla Juhász, Kim L. Chang, Ian Jameson, Peter Ralph, Michelle L. Colgrave

Research outputs 2022 to 2026

Algae-derived protein has immense potential to provide high-quality protein foods for the expanding human population. To meet its potential, a broad range of scientific tools are required to identify optimal algal strains from the hundreds of thousands available and identify ideal growing conditions for strains that produce high-quality protein with functional benefits. A research pipeline that includes proteomics can provide a deeper interpretation of microalgal composition and biochemistry in the pursuit of these goals. To date, proteomic investigations have largely focused on pathways that involve lipid production in selected microalgae species. Herein, we report the current state of microalgal proteome …

Proteome Changes Resulting From Malting In Hordein-Reduced Barley Lines, Mahya Bahmani, Angéla Juhász, Utpal Bos, Mitchell G. Nye-Wood, Malcolm Blundell, Crispin A. Howitt, Michelle L. Colgrave

Research outputs 2022 to 2026

Hordeum vulgare L., commonly known as barley, is primarily used for animal feed and malting. The major storage proteins in barley are hordeins, known triggers of celiac disease (CD). Here, sequential window acquisition of all theoretical mass spectra (SWATH)-MS proteomics was employed to investigate the proteome profile of grain and malt samples from the malting barley cultivar Sloop and single-, double-, and triple hordein-reduced lines bred in a Sloop background. Using a discovery proteomics approach, 2688 and 3034 proteins were detected from the grain and malt samples, respectively. By utilizing label-free relative quantitation through SWATH-MS, a total of 2654 proteins …

Multiomics Approach Captures Hepatic Metabolic Network Altered By Chronic Ethanol Administration, Isin Tuna Sakallioglu, Bridget Tripp, Jacy Kubik, Carol A. Casey, Paul G. Thomes, Robert Powers

Chemistry Department: Faculty Publications

Using a multiplatform and multiomics approach, we identified metabolites, lipids, proteins, and metabolic pathways that were altered in the liver after chronic ethanol administration. A functional enrichment analysis of the multiomics dataset revealed that rats treated with ethanol experienced an increase in hepatic fatty acyl content, which is consistent with an initial development of steatosis. The nuclear magnetic resonance spectroscopy (NMR) and liquid chromatography–mass spectrometry (LC-MS) metabolomics data revealed that the chronic ethanol exposure selectively modified toxic substances such as an increase in glucuronidation tyramine and benzoyl; and a depletion in cholesterol-conjugated glucuronides. Similarly, the lipidomics results revealed that ethanol …

Quantifying Protein Quality To Understand Protein Homeostasis, Hsien-Jung Lavender Lin

Theses and Dissertations

Proteins are the center of all biochemical reactions in living organisms. Proteins need to be present at the right time, in the right place, with the correct concentration and have the right shape to carry their designated function. Protein homeostasis is when all proteins in the proteome are in functional balance, and such balance is maintained by synthesis, folding, and degradation machinery. When protein homeostasis is lost, organisms start to age and develop diseases. To truly unveil disease mechanisms and provide more efficient means for treatment and prevention, we need a holistic understanding of the mechanism of protein homeostasis. Currently, …

Lipid Rafts, Exosomal Vesicles And Anti-Giardial Therapies, Brian Ivan Grajeda

Open Access Theses & Dissertations

Giardia lamblia, a protozoan parasite, is a major cause of waterborne infection, worldwide. While the trophozoite form of this parasite induces pathological symptoms in the gut, the cyst forms transmit the infection via contaminated water. Since Giardia is a non-invasive parasite, the actual mechanism by which it causes infection remains elusive. We have previously reported that Giardia assembles cholesterol and GM1 glycolipid-enriched lipid rafts (LRs) that participate in encystation and cyst production. To further delineate the role of LRs in pathogenesis, we isolated LRs from Giardia and subjected them to proteomic analysis. Various cellular proteins including the virulent proteins—e.g., giardins, …

Micro-Rna And Proteomic Profiles Of Plasma-Derived Exosomes From Irradiated Mice Reveal Molecular Changes Preventing Apoptosis In Neonatal Cerebellum, Simonetta Pazzaglia, Barbara Tanno, Ilaria De Stefano, Paola Giardullo, Simona Leonardi, Caterina Merla, Gabriele Babini, Seda Tuncay Cagatay, Ammar Mayah, Munira Kadhim, Fiona Lyng, Christine Von Toerne, Zohaib N. Khan, Prabal Subedi, Soile Tapio, Anna Saran, Mariateresa Mancuso

Articles

Cell communication via exosomes is capable of influencing cell fate in stress situations such as exposure to ionizing radiation. In vitro and in vivo studies have shown that exosomes might play a role in out-of-target radiation effects by carrying molecular signaling mediators of radiation damage, as well as opposite protective functions resulting in resistance to radiotherapy. However, a global understanding of exosomes and their radiation-induced regulation, especially within the context of an intact mammalian organism, has been lacking. In this in vivo study, we demonstrate that, compared to sham-irradiated (SI) mice, a distinct pattern of proteins and miRNAs is found …

Sd2 Ptov1 Interactomics, Joshua Andersen

ScholarsArchive Data

PTOV1 interactome data

Sd1 14-3-3 Interactome For Ptov1 Manuscript, Joshua Andersen

ScholarsArchive Data

14-3-3 interactome

Sd1 14-3-3 Interactome For Ptov1 Manuscript, Joshua Andersen

ScholarsArchive Data

Excel data sheet of MS data from 14-3-3 interactomics

Differential Proteomics Analysis Of Leaves From Hordeum Brevisublatum Under Salt Stress, Jinglin Shen, Xia Wu, Hongyu Tang, Jing Zhang, Hengtong Fang, Xudong Shi, Xue Bai, Zhuo Hao

IGC Proceedings (1997-2023)

No abstract provided.

Validation Of A Deployable Proteomic Assay For The Serological Screening Of Sexual Assault Samples, Catherine O'Sullivan Brown

Electronic Theses and Dissertations

Protein mass spectrometry (MS) has emerged as a technique to supplant traditional serological tests for body fluid identification. It was hypothesized that proteomic techniques would surpass the sensitivity and specificity of traditional serological techniques. An automated workflow coupled with protein MS has been developed for the confirmatory identification of five biological fluids. A developmental validation was completed, assessing parameters such as reproducibility, sensitivity, ion suppression, and limit of detection. Implementation was determined through tandem sample processing by MS, traditional serological tests, and standard DNA profiling methods. The MS approach offered superior detection limits while also providing true confirmatory results, producing …

Ensemble Protein Inference Evaluation, Kyle Lee Lucke

Graduate Student Theses, Dissertations, & Professional Papers

The Protein inference problem is becoming an increasingly important tool that aids in the characterization of complex proteomes and analysis of complex protein samples. In bottom-up shotgun proteomics experiments the metrics for evaluation (like AUC and calibration error) are based on an often imperfect target-decoy database. These metrics make the inherent assumption that all of the proteins in the target set are present in the sample being analyzed. In general, this is not the case, they are typically a mix of present and absent proteins. To objectively evaluate inference methods, protein standard datasets are used. These datasets are special in …

Proteome And Nutritional Shifts Observed In Hordein Double-Mutant Barley Lines, Utpal Bose, Angéla Juhász, Ronald Yu, Mahya Bahmani, Keren Byrne, Malcolm Blundell, James A. Broadbent, Crispin A. Howitt, Michelle L. Colgrave

Research outputs 2014 to 2021

Lysine is the most limiting essential amino acid in cereals, and efforts have been made over the decades to improve the nutritional quality of these grains by limiting storage protein accumulation and increasing lysine content, while maintaining desired agronomic traits. The single lys3 mutation in barley has been shown to significantly increase lysine content but also reduces grain size. Herein, the regulatory effect of the lys3 mutation that controls storage protein accumulation as well as a plethora of critically important processes in cereal seeds was investigated in double mutant barley lines. This was enabled through the generation of three hordein …

Proteome Analysis And Epitope Mapping In A Commercial Reduced-Gluten Wheat Product, Mitchell G. Nye-Wood, Angela Juhasz, Utpal Bos, Michelle L. Colgrave

Research outputs 2014 to 2021

Gluten related disorders, such as coeliac disease, wheat allergy and baker's asthma are triggered by proteins present in food products made from wheat and related cereal species. The only treatment of these medical illnesses is a strict gluten-free diet; however, gluten-free products that are currently available in the market can have lower nutritional quality and are more expensive than traditional gluten containing cereal products. These constraints have led to the development of gluten-free or gluten-reduced ingredients. In this vein, a non-GMO wheat flour that purports to contain “65% less allergenic gluten” was recently brought to market. The present study aims …

Novel Functional Proteomics Techniques For Multiplex And Single Cell Cytokines Detection, Mohammed Abdullah

Legacy Theses & Dissertations (2009 - 2024)

Since the early stages of biological research, the vital role of molecules in living creatures has been investigated and documented. According to Francis Watson's definition of the central dogma of biology, proteins are the final product of molecular biology and can't transfer information to other proteins like DNA or mRNA. Therefore, they are considered as the functional molecules of biology making them are more clinically relevant. However, the available techniques to study the protein characters, which coined laterally as proteomics science, are not well developed as genomics techniques did. Since the first human genome project is finished, the cost of …

Deep-Ultraviolet Laser Ablation For Biomolecular Analysis, Oluwaremilekun Omowunmi Lawal

LSU Doctoral Dissertations

New sampling and ionization techniques were developed using a deep-ultraviolet (DUV) laser at 193 nm wavelength for analysis of large biomolecules. A micrometer spatial resolution DUV laser ablation and capture system was constructed to sample tissue biomolecules. The spot size measurements of the focused beam showed that subcellular spots with ca. 2 μm diameter can be readily achieved with this wavelength. To demonstrate the extraction of tissue biomolecules with the DUV laser, regions of tissue sections were ablated and captured for offline proteomic and genomic analysis. The proteomic studies revealed that the tissue proteins were ejected without fragmentation and that …

Plasma And Serum Proteins Bound To Nanoceria: Insights Into Pathways By Which Nanoceria May Exert Its Beneficial And Deleterious Effects In Vivo, D. Allan Butterfield, Binghui Wang, Peng Wu, Sarita S. Hardas, Jason M. Unrine, Eric A. Grulke, Jian Cai, Jon B. Klein, William M. Pierce, Robert A. Yokel, Rukhsana Sultana

Chemistry Faculty Publications

Nanoceria (CeO2, cerium oxide nanoparticles) is proposed as a therapeutic for multiple disorders. In blood, nanoceria becomes protein-coated, changing its surface properties to yield a different presentation to cells. There is little information on the interaction of nanoceria with blood proteins. The current study is the first to report the proteomics identification of plasma and serum proteins adsorbed to nanoceria. The results identify a number of plasma and serum proteins interacting with nanoceria, proteins whose normal activities regulate numerous cell functions: antioxidant/detoxification, energy regulation, lipoproteins, signaling, complement, immune function, coagulation, iron homeostasis, proteolysis, inflammation, protein folding, protease inhibition, adhesion, protein/RNA …

Intrinsic Buffer Hydroxyl Radical Dosimetry For Hydroxyl Radical Protein Footprinting, Addison Roush

Honors Theses

Hydroxyl radical protein footprinting (HRPF) coupled to mass spectrometry is a powerful technique for the analysis of protein topography as it generates covalent mass labels that can survive downstream sample handling, and it is sensitive to the solvent accessibility of amino acid sidechains. Of the multiple platforms for HRPF, fast photochemical oxidation of proteins (FPOP) utilizes a pulsed 248 nm KrF excimer laser to label proteins by photolyzing hydrogen peroxide. FPOP is the most widely used HRPF platform because it labels proteins faster than unfolding can occur. Variations in FPOP sample conditions make it difficult to compare results between experiments …

Multi-Omics Strategies For Decoding Smoke-Assisted Germination Pathways And Seed Vigour, Utpal Bose, Angéla Juhász, James A. Broadbent, Setsuko Komatsu, Michelle L. Colgrave

Research outputs 2014 to 2021

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. The success of seed germination and the successful establishment of seedlings across diverse environmental conditions depends on seed vigour, which is of both economic and ecologic importance. The smoke-derived exogenous compound karrikins (KARs) and the endogenous plant hormone strigolactone (SL) are two classes of butanolide-containing molecules that follow highly similar signalling pathways to control diverse biological activities in plants. Unravelling the precise mode-of-action of these two classes of molecules in model species has been a key research objective. However, the specific and dynamic expression of biomolecules upon stimulation by these signalling …

Synthesis And Evaluation Of Novel Silica Hydride-Based Stationary Phases For Bioanalytical Applications

Seiichiro Watanabe