paegel Lab @ TSRI 130 Scripps Way 1B2 Jupiter, FL

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TOTAL SYNTHESIS OF A CELL

The controlled and rational synthesis of cellular supramolecular assemblies, such as bilayers and vesicles remains elusive. Our ability to deconstruct cells to investigate biological function far exceeds our capacity to construct and reconstitute biological activity in vitro. We are developing new microfluidic and chemical tools for synthesizing cellular envelopes and organelle-like scaffolds that afford control over encapsulation, membrane size and lamellarity. These constructs will enable detailed, mechanistic studies of complex biological activities, such as protein translocation and membrane insertion, proteolytic processing and protein modification, vesicular budding, endocytosis, and cell division, and will find applications in high-throughput screening reagent development and drug delivery.


MASS SPECTROMETRY, PROTEOMICS & PROTEASES

Mass spectrometry has ushered in a new era for proteomics. Powerful peptide sequencing and mapping technologies now allow us to glimpse the entire proteinaceous milieu of the living cell, its current state of chemical modification, and the correlation of protein expression and modification state with homeostasis and disease pathology. However our view is just a glimpse. Proteolytic digestion is the first step in protein analysis and it is limited often to trypsin, and regions of sequence lacking sufficient tryptic cleavage sites remain undetected. These sequence coverage gaps fragment our view of modification sites, which are central to metabolic function and diagnostic of dysregulation. We have developed a powerful compartmentalized evolution platform for discovering mutant proteases with orthogonal cleavage specificity. These new protease tools will enable complete protein sequence coverage and modification mapping using mass spectrometry.


MINIATURIZATION AND SMALL MOLECULE DISCOVERY

Conventional small molecule discovery is driven by high-throughput screening (HTS) centers, much like the center here on TSRI’s Florida campus. These centers are stuffed with expensive robotics designed to move microplates between various operations, such as compound dispensing, assay dispensing, incubation and optical screening. With well over 50,000 selective small molecule modulators required to probe human biology alone, our rate of discovery/cost (200 probes, 3 years, $400 million) must improve to reach this important goal. We are engineering an ultra-miniaturized and integrated microfluidic platform that dispenses with the robotics, microplates, and conventional compound libraries, and all of the costs associated therein. We envision an HTS instrument that can operate in any laboratory using DNA-encoded libraries, and consume minute quantities of precious screening reagents and cell lines. HTS campaigns of tomorrow will be as distributed and inexpensive as DNA sequencing is today.