A5043823252- Connexins and lens biology
- Mass Spectrometry Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Heat shock proteins research
- Metabolomics and Mass Spectrometry Studies
- Advanced Glycation End Products research
- Urinary Bladder and Prostate Research
- Ion Transport and Channel Regulation
- Enzyme Structure and Function
- Retinal Diseases and Treatments
- S100 Proteins and Annexins
- Biochemical effects in animals
- Helicobacter pylori-related gastroenterology studies
- Ion-surface interactions and analysis
- Retinal Development and Disorders
- Receptor Mechanisms and Signaling
- Aldose Reductase and Taurine
- Analytical Chemistry and Chromatography
- Protein Structure and Dynamics
- Polyamine Metabolism and Applications
- Analytical chemistry methods development
- Spectroscopy Techniques in Biomedical and Chemical Research
- Galectins and Cancer Biology
- Amino Acid Enzymes and Metabolism
- Ocular Surface and Contact Lens
Vanderbilt University
2016-2025
Vanderbilt University Medical Center
2009-2024
Nashville Oncology Associates
2012
Meharry Medical College
2012
Vanderbilt Health
2011
Medical University of South Carolina
2001-2010
Ralph H. Johnson VA Medical Center
2008
Mayo Clinic
2008
West Virginia University
2006
College of Charleston
2006
Abstract Oxidation is known to affect the structure, activity, and rate of degradation proteins, believed contribute a variety pathological conditions. Metal‐catalyzed oxidation (MCO) primary oxidizing system in many cell types. In this study, oxidative effects MCO (the Fenton reaction) on structure tryptophan residues α‐crystallin were determined. Tandem mass spectrometry (MS/MS) was utilized identify specific methionine products bovine sequence. After exposure, digested with trypsin,...
The role of vertebrate histone proteins or derived peptides as innate immune effectors has only recently been appreciated. In this study, high levels core H2A, H2B, H3 and H4 were found in hemocytes from the Pacific white shrimp, Litopenaeus vannamei . identified by in‐gel digestion, mass spectrometry analysis, homology searching. L. to be highly homologous histones other species. Based on homology, H2A was cloned its N‐terminus resemble known antimicrobial buforin I, parasin, hipposin....
Exosomes are membrane vesicles that secreted by cells upon fusion of multivesicular bodies with the plasma membrane. Exosomal proteomics has emerged as a powerful approach to understand molecular composition exosomes and potential accelerate biomarker discovery. Different proteomic analysis methods have been previously employed establish several exosome protein databases. In this study, TFE solution-phase digestion was compared in-gel found yield similar results. Proteomic urinary performed...
The accumulation of lipofuscin in the RPE is a hallmark aging eye. best characterized component A2E, bis-retinoid byproduct normal retinoid visual cycle, which exhibits broad spectrum cytotoxic effects vitro. purpose our study was to correlate distribution and A2E across human RPE.Lipofuscin fluorescence imaged flat-mounted from donors various ages. spatial distributions its oxides were determined using matrix-assisted laser desorption-ionization imaging mass spectrometry (MALDI-IMS) on...
Matrix assisted laser desorption ionization imaging mass spectrometry (MALDI IMS) has the ability to provide an enormous amount of information on abundances and spatial distributions molecules within biological tissues. The rapid progress in development this technology significantly improves our analyze smaller areas features mammalian eye evolved over millions years become essential asset for survival, providing important sensory input organism's surroundings. highly complex retina is...
Gastric carcinogenesis is mediated by complex interactions among Helicobacter pylori, host, and environmental factors. Here, we demonstrate that H. pylori augmented gastric injury in INS-GAS mice under iron-deficient conditions. Mechanistically, these phenotypes were not driven alterations the microbiota; however, discovery-based targeted metabolomics revealed bile acids significantly altered pylori–infected with iron deficiency, significant upregulation of deoxycholic acid (DCA), a...
Increased flux through the hexosamine biosynthesis pathway has been implicated in development of glucose-induced insulin resistance and may promote modification certain proteins with O-linked N-acetylglucosamine ( O-GlcNAc). L6 myotubes (a model skeletal muscle) were incubated for 18 h 5 or 25 mM glucose without 10 nM insulin. As assessed by immunoblotting an O-GlcNAc-specific antibody, high and/or enhanced O-GlcNAcylation numerous proteins, including transcription factor Sp1, a known...
A tissue preparation protocol for MALDI (matrix-assisted laser desorption/ionization) imaging mass spectrometry of integral membrane proteins was developed using ocular lens and retinal tissues as model samples. Frozen bovine human lenses were cryosectioned equatorially or axially at −20 °C into 20 μm-thick sections. Lens sections mounted onto gold-coated targets by methanol soft-landing to maintain integrity. Tissue underwent extensive water washing deplete the samples highly abundant...
To develop a protocol for MALDI (matrix-assisted laser desorption ionization) imaging mass spectrometry mapping the distributions of alpha-crystallin and its modified forms in human lens tissue as function age cataract.Frozen lenses were cryosectioned equatorially axially into 20-mum-thick sections, sections mounted onto conductive glass slides by methanol soft-landing. An ethanol washing procedure facilitated uniform matrix crystal formation two-step deposition to produce high-quality...
Summary Nonenzymatic post‐translational modification ( PTM ) of proteins is a fundamental molecular process aging. The combination various modifications and their accumulation with age not only affects function, but leads to crosslinking protein aggregation. In this study, aged human lens were examined using HPLC –tandem mass spectrometry blind search strategy. Multiple thioether S er T hr residues by glutathione GSH its metabolites unambiguously identified. Thirty‐four 36 sites identified...
Abstract Spatial proteomics has the potential to significantly advance our understanding of biology, physiology and medicine. Matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) is a powerful tool in spatial field, enabling direct detection registration protein abundance distribution across tissues. MALDI-MSI preserves histology allowing unbiased analysis complex, heterogeneous However, faces challenge simultaneous peptide quantification identification. To...
Because of the lack protein turnover in fiber cells ocular lens, Aquaporin 0 (AQP0), most abundant membrane undergoes extensive post-translational modification with cell age. To map distribution modified forms AQP0 within normal human lenses ranging age from 34 to 38 were concentrically dissected into several cortical and nuclear sections. Membrane proteins still embedded membranes digested trypsin, resulting C-terminal peptides analyzed by HPLC tandem mass spectrometry, permitting...
Aquaporin 0 (AQP0), the most abundant membrane protein in lens, is a water-permeable channel, has role fiber cell adhesion, and essential for structure organization. The purpose of this study was to identify proteins that interact with C terminus AQP0, by using proteomics approach, thus further elucidate AQP0 human lens.AQP0 C-terminal peptides antibody affinity chromatography were used purification interacting lens proteins. Purified digested trypsin, analyzed liquid (LC)-tandem mass...
We describe biological and experimental factors that induce variability in reporter ion peak areas obtained from iTRAQ experiments. demonstrate how these can be incorporated into a statistical model for use evaluating differential protein expression highlight the benefits of using analysis variance to quantify fold change. model's utility based on an data derived spike-in study.
MALDI imaging mass spectrometry (MALDI-IMS) has become a powerful tool for localizing both small molecules and intact proteins in wide variety of tissue samples normal diseased states. Identification imaged signals MALDI-IMS remains bottleneck the analysis limits interpretation underlying biology specimens. In this work, spatially directed microextraction followed by LC-MS/MS with electron transfer dissociation (ETD) was used to identify from specific locations three types; ocular lens,...
The human lens fiber cell insoluble membrane fraction contains important proteins, cytoskeletal and cytosolic proteins that are strongly associated with the membrane. purpose of this study was to characterize proteome phosphoproteome from lenses.HPLC-mass spectrometry-based multidimensional protein identification technology (MudPIT), without or phosphopeptide enrichment, applied membranes, respectively.In total, 951 were identified, including 379 integral membrane-associated proteins....
Urinary exosomes secreted by multiple cell types in the kidney may participate intercellular signaling and provide an enriched source of kidney-specific proteins for biomarker discovery. Factors that alter exosomal protein content remain unknown. To determine whether endogenous exogenous hormones modify urinary content, we analyzed samples from 14 mildly hypertensive patients a crossover study during high-sodium (HS, 160 mmol/d) diet low-sodium (LS, 20 to activate...
Background α Carboxyl terminus 1 (α CT 1) is a 25–amino acid therapeutic peptide incorporating the zonula occludens‐1 (ZO‐1)–binding domain of connexin 43 (Cx43) that currently in phase 3 clinical testing on chronic wounds. In mice, we reported reduced arrhythmias after cardiac injury, accompanied by increases protein kinase Cε phosphorylation Cx43 at serine 368. Herein, characterize detailed molecular mode action mitigating ischemia‐reperfusion injury. Methods and Results To study...