Date of Award

December 2015

Degree Type

Thesis

Degree Name

Master of Science

Department

Chemistry

First Advisor

David H. Petering

Committee Members

Arsenio Pacheco, Alexander Arnold

Abstract

Fluorescent sensors have been widely used as microscopic tools to image Zn2+ on a cellular level. Recently, it has been established that the sensors TSQ and Zinquin form adducts with Zn-proteins and image fractions of the Zn-proteome.1 Since TSQ and Zinquin bind specifically to many Zn-proteins, it is hypothesized that other metal binding ligands, both synthetic and natural, may also bind to the Zn-proteome. Biologically active 1,10-phenanthroline (Phen) and related molecules were investigated for their ability to bind to Zn-proteome. Similarly, the cellular tripeptide, glutathione was investigated. It was observed that Phen and some other metal binding, bidentate ligands, were able to displace TSQ from the TSQ-Zn-proteome. Specifically when cellular Zn-proteome reacted with TSQ, adduct formation occurs, that was detected by fluorescence emission at 470 nm. Upon reaction of TSQ-Zn-proteome with 1,10-phenanthroline, glutathione, and some other related small molecules, a significant decrease in fluorescence was observed, indicating the displacement of TSQ by each competing ligand. Native SDS (NDS) PAGE electrophoresis of the product of the reaction of 1,10-Phenanthroline with TSQ-Zn-proteome further confirmed the displacement of TSQ from an array of Zn-proteins. This reaction was further characterized with the use of Zn-alcohol dehydrogenase as a model enzyme that reacts with TSQ and Phen. These studies reveal the Zn-proteome as widely available for adduct formation at its zinc binding sites. The importance of such reactions is being investigated.

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