Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 22 for “"Metabolic Labeling"”.
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Investigating sphingolipid behavior and function using metabolic labeling
… this complexity. Chapters 2 and 3 present metabolic labeling with fluorophores and stable isotope tags, respectively, as tools to investigate sphingolipid behaviors. Metabolic labeling enables one to detect and directly observe sphingolipids, and not the activities or levels of the enzymes …
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Selective in vivo metabolic labeling for cell targeting and immune regulation
Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2025-08-01
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Developing a Novel Cell Surface RNA Detecting and Profiling Method via RNA Metabolic Labeling
… and sensitivity . In this study, we use metabolic labeling and chemical cross-linking techniques to specifically label csRNA with biotin handles. We intended to use this technique for separating biotin-labeled csRNA from total RNA samples for characterization purposes. The primary …
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Dynast: Inclusive and efficient quantification of metabolically labeled transcripts in single cells
… biased approximations. Recent advancements in metabolic labeling enabled the direct, unbiased measurement of nascent RNA, yielding significantly improved RNA velocity estimates. However, there is still a lack of a standardized computational framework to process these data. This study introduces …
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Interrogation of Yeast Responses to Anaerobic Growth Conditions at the Intact Protein Level Using Improved Top-Down Proteomics
… development of the front-end platform and metabolic labeling quantitative strategy, top-down proteomics was employed to investigate the response of Saccharomyces cerevisiae towards anaerobiosis. Approximately 150 protein pairs were measured with 4 differential expressions determined …
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Models of dynamic RNA regulation in mammalian cells
… RNA production rates by coupling short RNA metabolic labeling with advanced RNA quantification. I leverage recent improvements in RNA quantification by next-generation sequencing technology, to significantly increase the resolution of metabolic labeling in both time and gene-structure. …
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Quantitative Proteomic Methodology Use and Development to Characterize Ethanol Modulation of Microglial Function
… in neuroinflammation. We use mass spectrometric metabolic labeling techniques to explore and quantify the microglial proteome in immortalized cell lines and in vivo enriched microglia. Our proteomic profiling and subsequent validation suggests that microglia do activate in response to ethanol …
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A Phosphoinositide-Specific Phospholipase C of Trypanosoma Cruzi: Unique Characteristics of a Delta-Type Homologue
… is lipid modified in vivo as demonstrated by metabolic labeling with [3H]myristate and [3H]palmitate, and immunoprecipitation with anti-TcPI-PLC antibody, and may constitute the first example of a new group of PI-PLCs.
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Stress response of porphyromonas gingivalis
… polyacrylamide gel electrophoresis after metabolic labeling with (14C)-amino acids. When P. gingivalis cells were shifted from 37°C to 42°C, elevated synthesis of four proteins with the apparent molecular weight of 92, 80, 74, and 62kDa was observed, whereas synthesis of a 50kDa protein …
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The development and application of mass-spectrometry-based tools to monitor proteome remodeling in microbes
… power of quantitative proteomics coupled with metabolic labeling to investigate how microbes remodel their proteome during cellular adaptation. In chapter 2, I describe the development and characterization of these proteomic methods, including a detailed analysis of the variety of metabolic …
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Understanding the dynamics of rhythmic gene expression in mammalian cells
… RNA dynamics to rhythmic gene expression, I used metabolic labeling and a novel computational pipeline to analyze transcriptomic data in synchronized NIH3T3 cells. I identified 685 rhythmically expressed RNAs with a period of 24-hour in my dataset, of those 389 were rhythmically synthesized and 24 …
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Nucleolar Stress Due to Depletion of Nopp140 in Drosophila melanogaster
… by RNAi. We used TEM, immuno-blot analysis, and metabolic protein labeling to show the loss of ribosomes. Targeted loss of Nopp140 only in larval wing discs caused apoptosis, which eventually led to defects in the adult wings. These defects were not rescued by a p53 gene deletion, as the …
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Regulation of Connexin43 by Phosphorylation in Heart Development
… principally high molecular weight forms of Cx43. Metabolic labeling procedures confirmed that these forms of Cx43 were phosphorylated, suggesting that the development of a functional, four-chambered heart requires gap junctional communication provided by Cx43 during embryogenesis which is …
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Development of a chemical biology approach for glycoprotein discovery in Caenorhabditis elegans reveals glycosylated isoforms of multiple proteins with mitochondrial function
… elegans primary embryonic cells for azido-sugar metabolic labeling and then used this method to identify novel C. elegans glycoprotein candidates (Chapter 2). Additionally, in a project led by another student, Dr. Casey Snodgrass, and in which I played a supporting role, our lab adapted this …
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Plasticity of the Nuclear Pore Complex Revealed With Proteomics
… multiple methodologies: affinity capture, metabolic labeling and mass spectrometry. Moreover, to identify the required affinity capture conditions I have developed a high-throughput screen, which has successfully worked with numerous protein complexes, in addition to the NPC, to reveal …
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Molecular Characterization of Dopamine Transporter Phosphorylation and Regulation
… in rDAT expressing LLC-PK1 cells by using 32PO4 metabolic labeling and [3H] DA uptake assays. Pretreatment with cocaine or methylphenidate (MPH) had no effect on basal or PMA stimulated DAT phosphorylation and DA transport. GBR 12909 suppressed PMA-induced DAT phosphorylation and internalization. …
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Characterization of Metastasis-Associated Cell Surface Glycoproteins in Prostate Cancer
… successfully developed, which combined selective metabolic labeling of cell surface sialyl glycans, chemically probing the labeled sugar with a biotin tag, affinity purification of sialylated proteins, SDS-PAGE separation, and subsequent LC-MS/MS for protein identification. Application of this …
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Role of O-GlcNAc in the Vertebrate Secretory Pathway
… Our approach utilizes both stable isotope labeling with amino acids in cell culture (SILAC) for quantitation and metabolic labeling of O-GlcNAc for enrichment. Using our glycoproteomics workflow we examined the effects of brefeldin A (BFA), a fungal metabolite that disrupts vesicle …
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MS-Based Chemical Proteomics Studies of Extracellular Glycoproteins: Identification, Quantification, and Dynamics
… proteomics. First, an enrichment method based on metabolic labeling with a sugar analog, copper-free click chemistry, and MS-based proteomics is employed to target cell-surface glycoproteins from multiple cell lines. The method is universal for all glycan types, targets surface glycoproteins only …
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A multiplexed approach for quantitative profiling of the translatome using bioorthogonal non-canonical amino acids
… Aha. By coupling pulsed stable isotope labeling of amino acids in cell culture (pSILAC) and Aha metabolic labeling of newly synthesized proteins with strain-promoted azide-alkyne cycloaddition and tandem mass tag (TMT) labeling, I am able to quantitatively interrogate the translatome in …
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