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 30 for “"Protein Labeling"”.
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Site-specific protein labeling via sortase A and its applications/
… purposes have ensured that methods for labeling of proteins with reporter molecules remain in high demand. Standard chemical labeling methods using entities that react with amino acid side chains lack the specificity to ensure precise placement of reporter groups. Genetic methods, …
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Molecular engineering of new protein labeling methodology based on rational design and in vitro evolution
Site-specific labeling using E coli biotin ligase (BirA) and its 15-amino acid "acceptor peptide" (AP) has been applied to study the function of various cellular proteins. In order to extend the capabilities of biotin ligase-based labeling, we engineered key elements of the labeling platform. First …
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Rational design and directed evolution of probe ligases for site-specific protein labeling and live-cell imaging
… superior photophysical properties to fluorescent proteins and are much smaller. However, in order to use these probes for live-cell protein imaging, highly specific labeling methods are required. Here, we will describe three efforts to re-engineer the E. coli enzyme, lipoic acid ligase (LplA), to …
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Methodology development for imaging histone modifications and for site-specific protein labeling in vitro and on the surface of living cells
… (FRET). These reporters are four-part chimeric proteins with the domains from N- to C-terminus as follows: a cyan fluorescent protein (CFP), a phosphoserine recognition domain (14-3-3 tau) or a methyllysine recognition domain (HP1 or Pc chromodomain), a peptide whose sequence corresponds to the …
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Trimethoprim-Based Chemical Tags for High Resolution Live Cell Imaging
… developed to complement the use of fluorescent proteins in live cell imaging. Chemical tags retain the specificity of protein labeling achieved with fluorescent proteins through genetic encoding, but provide smaller, more robust tags and modular use of organic fluorophores with high …
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Site-specific labeling of cellular proteins with unnatural substrates of biotin ligases
… develop a general methodology for site-specific protein labeling with biophysical probes. We synthesized an isosteric ketone analog of biotin and discovered that BirA also ligates it to the AP with similar kinetics while retaining the high peptide specificity of the native reaction. Because …
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Characterization of protein induced fluorescence enhancement (PIFE): A label-free protein assay with short distance sensitivity
… used for monitoring the interactions between a protein and a nucleic acid. It typically requires a site-specific labeling of the protein, which can be labor-intensive and inefficient. In addition, for common dye pairs, the distance sensitivity of FRET lies only between 3-7 nm. In this report, we …
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Enzyme-based reporters for mapping proteome and imaging proteins in living cells
… functions are uniquely defined by the set of proteins they possess. For each individual protein, precise targeting to a specific sub-cellular location and trafficking between compartments are often key to its proper function. In fact, many human diseases are linked to mutations that cause …
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Chemical and enzymatic tools to study proteins in their native cellular environment
… trafficking, and turnover of individual proteins, strategies that allow the tagging, and imaging, and identification of entire proteomes have also offered valuable insights into disease biology. Since protein visualization serves as a complement to protein identification, this thesis …
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Chemical biology of matrix metalloproteases
… biological functions through activity-based protein labeling. Herein we introduce synthetic routes toward different P1a?? substituted MMPI warheads and their use in the MMPI library synthesis on solid phase. In addition, by taking the advantages of the relative ease and convenience of …
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NEW METHODS FOR DETECTING BIOMARKERS OF OXIDATIVE STRESS AND REDOX SIGNALING ON PROTEIN CYSTEINE RESIDUES
… and formation of these species reversibly alters protein function. Protein oxidation is studied to a lesser extent than lipid and DNA oxidation in part because of a lack of sensitive, stable, readily detectable markers for tracking these unstable intermediates.
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METHOD DEVELOPMENT FOR THE ANALYSIS OF PROTEINS AND SMALL
… in order to be detected via LIF. Furthermore, protein separations using CE-LIF may be complicated by protein interactions with the negatively charged surface of the inner capillary wall, arising from the ionization of surface silanol groups. Presented here is a method for quantitation of …
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Identifying Sortase A Variants With Higher Catalytic Effeciency
<p>In the past two decades, the field of protein engineering has evolved rapidly to include new genetic and chemical techniques to alter protein function. Protein engineering seeks to improve enzyme properties through powerful methods that specifically incorporate novel or improved function in …
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COMPARISON OF E. COLI AND Y. PESTIS LPS TLR4 TIME COURSE ON P388D1 MACROPHAGES USING FLUORESCENT CONJUGATED ANTIBODIES
… cellular response to infectious agents, and protein interactions are essential for this response. However, the protein interactions involved in cellular response to pathogens are not completely understood. Clarifying the manner in which proteins bind and respond to infectious agents is …
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Engineered heme peroxidases as genetically encoded probes with diverse applications in cell biology
Proteins perform myriad functions inside cells. In vitro studies of purified proteins yield important information about their function, but it is an important pursuit in cell biology to study proteins within the native context of intact cells, since their physiological functions might not be …
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Cellular delivery and site-specific targeting of organic fluorophores for super-resolution imaging in living cells
… encoded probes to specific cellular proteins with high specificity and minimal interference. We describe herein the development of new methods for cellular delivery and sitespecific targeting of organic fluorophores to proteins in living cells. Building on our lab's previous work on …
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Nucleolar Stress Due to Depletion of Nopp140 in Drosophila melanogaster
… of the related nucleolar and Cajal body phospho-protein, Nopp140, in Drosophila melanogaster led to nucleolar stress and eventual lethality when multiple tissues were depleted of Nopp140 by RNAi. We used TEM, immuno-blot analysis, and metabolic protein labeling to show the loss of ribosomes. …
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Microfluidic-based Fluorescence Detection for Biomarker Analysis
… flow platform was applied to enable surface protein profiling on exosomes. The platform was validated with single molecules first, including alexa647 anti-IgG and PE anti-IgG. The recovery rate for these 2 molecules were higher than 90%, indicating the high sensitivity of detecting single …
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