Design and synthesis of probes for detection of protein-protein interaction and RNA localization

Abstract

The use of the ketone biotin - benzophenone-biotin hydrazide system for detecting the formation of cyan fluorescent protein and NF-kappaB p50 dimers was assessed. A series of benzophenone-based probes were synthesized and tested for photocrosslinking activity to investigate the efficiency of photocrosslinking in these systems. Three series of small molecule probes were synthesized for the selection of ribozymes from a random sequence pool. Solid-phase immobilized fluorescein and fluorescein phosphates were synthesized for the indirect selection of a fluorescein phosphatase ribozyme. A corresponding thiophosphate analog was created for the in-gel selection of a thiophosphatase ribozyme via APM-PAGE. Finally, a series of fluorescein-nucleoside phosphate conjugates was designed and synthesized for use in the solution phase preparation of a fluorogenic ribozyme substrate, and later immobilization of this substrate on a silyl resin for direct ribozyme selection.