A5087464233- Advanced Proteomics Techniques and Applications
- Mass Spectrometry Techniques and Applications
- RNA and protein synthesis mechanisms
- Bacterial biofilms and quorum sensing
- Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Autophagy in Disease and Therapy
- Tissue Engineering and Regenerative Medicine
- Metabolomics and Mass Spectrometry Studies
- Biosensors and Analytical Detection
- Legionella and Acanthamoeba research
- Analytical Chemistry and Chromatography
- Pluripotent Stem Cells Research
- Doctoral Education Challenges and Solutions
- Cancer-related molecular mechanisms research
- Molecular Biology Techniques and Applications
- Identification and Quantification in Food
- Muscle metabolism and nutrition
- Muscle Physiology and Disorders
- PARP inhibition in cancer therapy
- Ion-surface interactions and analysis
- Perfectionism, Procrastination, Anxiety Studies
- Cardiac electrophysiology and arrhythmias
- Advanced Biosensing Techniques and Applications
- Enzyme Structure and Function
- Vibrio bacteria research studies
University of Wisconsin–Madison
2016-2025
National Institute of Allergy and Infectious Diseases
2020-2024
National Institutes of Health
2020-2024
Government of the United States of America
2023
Mass spectrometry (MS)-based top-down proteomics is a powerful method for the comprehensive analysis of proteoforms that arise from genetic variations and post-translational modifications (PTMs). However, MS high molecular weight (MW) proteins remains challenging mainly due to exponential decay signal-to-noise ratio with increasing MW. Size exclusion chromatography (SEC) favored size-based separation biomacromolecules but typically suffers low resolution. Herein, we developed serial size...
Significance Hypertrophic cardiomyopathy (HCM) is a common genetic heart disease and leading cause of sudden cardiac death in young adults. HCM has been linked to mutations genes encoding sarcomeric proteins, but how different can result similar clinical phenotype unknown. Analysis surgical tissue samples from patients with severe outflow track obstruction using high-resolution mass spectrometry–based top-down proteomics revealed pattern altered proteoforms across tissues compared...
Human pluripotent stem cell (hPSC)-derived cardiomyocytes exhibit the properties of fetal cardiomyocytes, which limits their applications. Various methods have been used to promote maturation hPSC-cardiomyocytes; however, there is a lack an unbiased and comprehensive method for accurate assessment maturity hPSC-cardiomyocytes.
Mass spectrometry (MS) based top-down proteomics provides rich information about proteoforms arising from combinatorial amino acid sequence variations and post-translational modifications (PTMs). Fourier transform ion cyclotron resonance (FT-ICR) MS affords ultrahigh resolving power high-accuracy mass measurements, presenting a powerful tool for characterization of proteoforms. However, the detection large proteins complex mixtures remain challenging due to exponential decrease in S:N with...
Mass spectrometry (MS)-based proteomics provides unprecedented opportunities for understanding the structure and function of proteins in complex biological systems; however, protein solubility sample preparation before MS remain a bottleneck preventing high-throughput proteomics. Herein, we report bottom-up proteomic method enabled by newly developed MS-compatible photocleavable surfactant, 4-hexylphenylazosulfonate (Azo) that facilitates robust extraction, rapid enzymatic digestion (30 min...
Although top-down proteomics has emerged as a powerful strategy to characterize proteins in biological systems, the analysis of endogenous membrane remains challenging due their low solubility, abundance, and complexity subproteome. Here, we report simple but effective enrichment separation for enabled by cloud point extraction multidimensional liquid chromatography coupled high-resolution mass spectrometry (MS). The efficiently enriched using single extraction, eliminating need...
Defining the proteoform landscape of breast cancer can provide unique insights into signaling pathways driving disease progression. While bottom-up proteomics has been utilized to profile cancer, it lacks ability capture intact proteoforms that may underpin disease. Top-down is ideally suited characterize proteoforms; however, most top-down studies have limited low molecular weight (MW) proteins (<50 kDa). Herein, we employed a two-dimensional (2D) liquid chromatography combining size...
Determining changes in protein expression and post-translational modifications (PTMs) is crucial for elucidating cellular signal transduction disease mechanisms. Conventional antibody-based approaches have inherent problems such as the limited availability of high-quality antibodies batch-to-batch variation. Top-down mass spectrometry (MS)-based proteomics has emerged most powerful method characterization quantification modifications. Nevertheless, robust methods to simultaneously determine...
Top-down mass spectrometry (MS)-based proteomics has become a powerful tool for comprehensive characterization of intact proteins. However, because the high complexity proteome, highly effective separation proteins from complex mixtures prior to MS analysis remains challenging. Monolithic columns have shown great promise protein due their permeability, low backpressure, and fast transfer. Herein, first time, we developed bridged hybrid bis(triethoxysilyl)ethylene (BTSEY) monolith with C8...
Bacteria can use chemical signals to assess their local population density in a process called quorum sensing (QS). Many of these bacteria are common pathogens, including Gram-positive that utilize agr QS systems regulated by macrocyclic autoinducing peptide (AIP) signals. Listeria monocytogenes, an important foodborne pathogen, uses system regulate variety virulence factors and biofilm formation, yet little is known about the specific roles infection its persistence various environments....
Three-dimensional (3D) human induced pluripotent stem cell-derived engineered cardiac tissues (hiPSC-ECTs) have emerged as a promising alternative to two-dimensional hiPSC-cardiomyocyte monolayer systems because hiPSC-ECTs are closer representation of endogenous and more faithfully reflect the relevant pathophysiology. The ability perform functional molecular assessments using same hiPSC-ECT construct would allow for reliable correlation between observed performance underlying events, thus...
Enterohemorrhagic Escherichia coli (EHEC) is the causative agent of severe diarrheal disease in humans. Cattle are natural reservoir EHEC, and approximately 75% EHEC infections humans stem from bovine products. Many common bacterial pathogens, including rely on chemical communication systems, such as quorum sensing (QS), to regulate virulence facilitate host colonization. uses SdiA E. (SdiAEC), an orphan LuxR-type receptor, sense N-acyl l-homoserine lactone (AHL) QS signals produced by other...
We have used mass spectrometry (MS) to characterize protein signaling in lipopolysaccharide (LPS)-stimulated macrophages from human blood, THP1 cells, mouse bone marrow, and Raw264.7 cells. Protein ADP-ribosylation was truncated down phosphoribose, allowing for enrichment identification of the resulting phosphoribosylated peptides alongside phosphopeptides. Size exclusion chromatography-MS (SEC-MS) separate proteoforms by size; complexes were then identified weighted correlation network...
Heterogeneity in skeletal muscle contraction time, peak power output, and resistance to fatigue, among others, is necessary accommodate the wide range of functional demands imposed on body. Underlying this heterogeneity are a myriad differences myofilament protein isoform expression post-translational modifications; yet, characterizing remains challenging. Herein, we have utilized top-down liquid chromatography (LC)–mass spectrometry (MS)-based proteomics characterize proteoform seven rat...
Proteoforms, the primary effectors of biological processes, are different forms proteins that arise from molecular processing events such as alternative splicing and post-translational modifications. Heart diseases exhibit changes in proteoform levels, motivating development a deeper understanding heart landscape. Our recently developed two-dimensional top-down proteomics platform coupling serial size exclusion chromatography (sSEC) to reversed-phase (RPC) expanded coverage human proteome...
Abstract Mass spectrometry (MS)‐based proteomics provides unprecedented opportunities for understanding the structure and function of proteins in complex biological systems; however, protein solubility sample preparation before MS remain a bottleneck preventing high‐throughput proteomics. Herein, we report bottom‐up proteomic method enabled by newly developed MS‐compatible photocleavable surfactant, 4‐hexylphenylazosulfonate (Azo) that facilitates robust extraction, rapid enzymatic digestion...
Electron capture dissociation (ECD) is well suited for the characterization of phosphoproteins, with which labile phosphate groups are generally preserved during fragmentation process. However, impact phosphorylation on ECD intact proteins remains unclear. Here, we have performed a systematic investigation effect by comparing cleavages mono-phosphorylated α-casein, multi-phosphorylated β-casein, and immunoaffinity-purified phosphorylated cardiac troponin I those their unphosphorylated...
This article describes a graduate student-led effort to develop climate survey assess, advocate for, and improve the well-being mental health of students postdocs in Department Chemistry at University Wisconsin–Madison. Graduate have an increased incidence depression relative general population, given transient nature student understanding addressing concerns can be challenging. The goal this is illustrate how students, with support departmental faculty, staff, existing on-campus resources,...