Development of Arginyl-tRNA synthetase from Pseudomonas aeruginosa as a platform to screen for inhibitors of protein synthesis

Author

Daniel Cantu, The University of Texas Rio Grande Valley

Date of Award

5-2016

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

First Advisor

Dr. James Bullard

Second Advisor

Dr. Evangelia Kotsikorou

Third Advisor

Dr. Megan Keniry

Abstract

Pseudomonas aeruginosa is a Gram- negative bacteria that is the leading cause of mortality in immunocompromised patients. Arginyl-tRNA synthetase (ArgRS) is an enzyme that catalyzes the covalent attachment of arginine to the cognate tRNAArg during protein synthesis. Scintillation proximity assays (SPA) were adapted to the aminoacylation assay and used to screen, one natural compound library (800) and one synthetic compound library (890) to identify inhibitors of the activity of ArgRS. Five compounds were identified which inhibited greater than 50% of enzymatic activity, two of these compounds (BT04F10 and BT11F03) exhibited promising MICs against Gram+ bacteria and moderate activity against Gram- bacteria. These two compounds were analyzed for mechanism-of-action against Haemophilus influenzae and Staphylococcus aureus in time-kill kinetics and both were observed to be bacteriostatic. The two compounds were tested to determine mechanism of action relative to the natural substrates, ATP and arginine, and found to be non-competitive with both substrates.

Comments

Copyright 2016 Daniel Cantu. All Rights Reserved.

https://www.proquest.com/dissertations-theses/development-arginyl-trna-synthetase-i-pseudomonas/docview/1810996018/se-2?accountid=7119

Recommended Citation

Cantu, Daniel, "Development of Arginyl-tRNA synthetase from Pseudomonas aeruginosa as a platform to screen for inhibitors of protein synthesis" (2016). Theses and Dissertations. 15.
https://scholarworks.utrgv.edu/etd/15