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 9 of 9 for “"site-specific labeling"”.
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Site-specific labeling of cellular proteins with unnatural substrates of biotin ligases
E. coli biotin ligase (BirA) catalyzes the site-specific ligation of biotin to the lysine within its 15-amino acid peptide substrate (AP). We harnessed the high peptide substrate specificity of BirA to develop a general methodology for site-specific protein labeling with biophysical probes. We …
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Characterization of protein induced fluorescence enhancement (PIFE): A label-free protein assay with short distance sensitivity
… 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 report on a newly developed fluorescence assay …
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DNA-catalyzed covalent modification of amino acid side chains in tethered and free peptide substrates
… reductive amination that may be applicable to site-specific labeling of nucleic acids. Finally, the thesis summarizes the optimization of the incubation conditions using click chemistry for direct free peptide selection. From these studies, we gained insights on how to direct our future …
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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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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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Chemical Tools for Exploring IFITM3 S-Palmitoylation and Mechanism
… been challenging but recent advances in chemical labeling methods have enabled more sensitive detection and proteomic analyses, which I summarize in Chapter 1. The proteomic analysis of S-palmitoylated proteins in dendritic cells and macrophages by our laboratory revealed that the …
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Protein engineering design principles for the development of biosensors
… We observe that analytes can harbor specific binding pockets for the fluorophore, which sharply increase the fluorescence produced upon binding. Furthermore, we demonstrate that a fluorophore conjugated to locally rigid surfaces possesses lower background fluorescence. Based on these …
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Enzyme-mediated labeling of proteins and protein-protein interactions in vitro and in living cells
… previously used by the Ting research group for site-specific labeling of peptide-tagged cell surface proteins. We sought to expand the utility of biotin ligase-mediated labeling to functional group handles, including azides and alkynes, for bio-orthogonal chemistry. Since the BirA and its point …
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Site-Specific Incorporation of Biochemical and Biophysical Probes into Proteins Using Expressed Protein Ligation
… a general strategy was developed for the site-specific incorporation of fluorophores into proteins using Abl-SH3 as a model system. In the second application, chemistries were developed that allowed the site-specific introduction of phospho-amino acids into proteins, in this case using the …