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Showing 1 to 12 of 12 for “"Kinetic assay"”.
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Biochemical Analysis of Poly(ethylene terephthalate) Film Degradation Kinetics of Engineered IsPETase Variants
… may not be directly applicable to kinetics of these enzymes, as the catalysis is occurring under a heterogeneous phase. To improve the fundamental understanding of the enzymatic reaction on the surface and derive an appropriate biochemical model for kinetic analysis, this thesis …
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Suitability of RelBE and MazEF toxin-antitoxin systems as antibacterial targets
… against the RelB antitoxin. Additionally, a kinetic assay using a fluorogenic substrate was designed for the ribonuclease toxin MazFSa and applied in a screen for peptide enhancers of MazFSa enzymatic activity. A highly sensitive radiometric gel-based assay allowed for the first detection of …
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On the Kinetics of Protein Misfolding and Aggregation
… it is demonstrated, that highly accurate kinetic experiments, using a novel biosensing method, can yield fundamental insight into the dynamics of proteins in the region of the free energy landscape corresponding to protein aggregation. First, a section on Method development describes the …
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The Role of Metabolism and Epigenetics in Regulating Cellular Plasticity
… mark in DNA. Using an in vitro real-time kinetic assay, we identified that one of the best substrates for three of the four human glycosylases examined is 5-carboxyuracil (5caU). 5caU is the least likely product to form as it requires several sequential oxidations and deamination reactions …
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Structure and function of enzyme I of the PTS
… enzyme I binding site on HPr was determined by a kinetic assay, using site-directed mutants of HPr as substrates for enzyme I. This site of interaction agreed very well with that found in the NMR solution structure of the complex of HPr with the N-terminal domain of enzyme I (Garrett 'et al'., …
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Expression, Purification, and Characterization of the Mast Cell Proteases Chymase and Cathepsin G.
… the highest levels of activity were selected by kinetic assay. Active chymase was purified from the culture media with a 22% yield of activity by a simple two-step procedure that involved hydrophobic-interaction chromatography followed by affinity chromatography on immobilized heparin. The major …
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Drug discovery for misfolding diseases using structure-based iterative learning
… for aggregation inhibition were obtained by an assay specifically isolating secondary nucleation, the major mechanism of toxic oligomer production. My results demonstrate that this approach leads to the facile identification of compounds which are two orders of magnitude more potent than …
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Structural and Functional Studies of Mycobacterial General Transcription Factors RbpA and CarD
… decades, the vast majority of biochemical and kinetic studies have used Escherichia coli (Eco) RNA polymerase (RNAP) as a model for studying prokaryotic transcription initiation. However, properties of Eco RNAP are not representative of RNAPs from other bacterial species. Transcription in …
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Investigation of Narrow Spectrum Targets in Antibacterial Drug Discovery
… including structural studies, biochemical assay development and application, and inhibitor screening and testing. Methods: An assortment of drug discovery techniques were employed for the targeting of SmTopoI and CdFabK, including different protein expression and purification techniques; …
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Mining biological diversity to optimize chlorite dismutase for water treatment
… homologs from a variety of bacterial species. A kinetic assay was developed to track product formation using a chloride ion selective electrode. This assay was used to quantify and compare the kinetics, shelf life, and catalytic life of the seven homologs. Additionally, a custom polyclonal …
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Split Inteins: From Mechanistic Studies to Novel Protein Engineering Technologies
… were obtained by the development of a novel kinetic assay that allowed for quantitative observation of a crucial intermediate in the protein splicing pathway, the branched intermediate (BI). Using this assay, BI resolution was unambiguously identified as the rate limiting step for Npu …