DEVELOPING HIGH-THROUGHPUT APPROACHES FOR ENZYME SCREENING AND PROTEOMIC PROFILING

Abstract

Enzymes play important roles in the functioning of any living system and regulating cellular processes. There are around 18-29% of eukaryotic genomes which encode enzymes. However, due to lack of efficient technologies, little is still understood about physiological roles, substrate specificity and downstream targets of these enzymes as well as what specific proteins are associated with cancer states, which are called biomarkers. This thesis examines and addresses these challenges by integration of microarrays, microplates and other enabling platforms. Chapter 2 and 3 respectively present an affinity and activity based microarray approach for mapping the substrate specificity of PTPs. With these approaches, potent and specific substrates could be discovered as well as new phosphatase/substrate pairs. Chapter 4 demonstrates an array based systematic approach for high-throughput discovery of potent and selective SH2 domains binders. Chapter 5 presents a systematic microarray approach for comparative proteomic profiling of mammlian cell lysates and biomarker discovery.