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Title: High-throughput methodologies for enzyme inhibitor profiling
Authors: WU HAO
Keywords: Small Molecule Microarray, High Throughput Screening, 14-3-3 Protein, Cysteine Proteases, Inhibitor, Protein Protein Interaction
Issue Date: 7-Mar-2011
Citation: WU HAO (2011-03-07). High-throughput methodologies for enzyme inhibitor profiling. ScholarBank@NUS Repository.
Abstract: Recent evidence suggests that 18~29% of eukaryotic genomes encode enzymes which are critical to the vital functioning of living system. However, only a limited proportion of these enzymes have thus far been studied, characterized, and little is understood about the physiological roles, substrate specificity and downstream targets of the vast majority of these important proteins. A key step towards the biological characterization of enzymes, as well as in their adoption as drug targets, is the development of global solutions that bridge the gap in understanding proteins and their interactions, especially in complex environments. This thesis examines and addresses these challenges by integration a series of technologies that span various analytical modes, effectively expanding current capabilities in protein profiling by leveraging on throughput. These include small molecule microarray (Chapter 2 and 3) and microplate (Chapters 4 & 5) platforms. Chapter 2 describes a novel peptide aldehyde microarray for rapid differentiation of infection stages in cellular level, and characterized the potential enzyme targets of identified compounds. Chapter 3 presents a novel hybrid small molecule library that enables the development of inhibitor targeting 14-3-3 protein interaction in vivo. Chapter 4 shows a robust screening and analysis of inhibitor library against various protein tyrosine phosphatase. Chapter 5 demonstrates an oriented library designed targeting caspases. Cohesively, the approaches are applied (but not limited) to investigations of certain classes of enzymes or proteins, but also provide functional insight into complex biological dynamics that orchestrate the remarkable enigma of life.
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