Abstract

M. tuberculosis contains around a dozen potential Nudix hydrolases, and we are characterizing several of these enzymes as potential novel antibiotic targets. The diadenosine polyphosphatases (ApnAases) / mRNA decapping enzymes are a family of enzymes within the Nudix hydrolase superfamily. In M. tuberculosis there is a primary Nudix ApnAase and a secondary Nudix ApnAase. The diadenosine polyphosphatases from Legionella pneumophilia and Bartonella bacilliformis have been found to be important in each pathogen's ability to invade human host cells. We are interested in whether these enzymes act in the same way in M. tuberculosis. If they are found to be involved in invasiveness and thus in virulence, then these enzymes could be novel antibiotic targets. We have cloned, overexpressed, and purified the primary Nudix ApnAase from M. tuberculosis, and solved its structure with molecular replacement. The diffraction data had a resolution of 2.53 Å and a completeness of 99.92% resulting in a refined model with an R-free of 0.2835 and an R-work of 0.2293. We were able to crystallize the primary ApnAase with ATP in the Nudix hydrolase active site, which provided insight into ten residues that are potentially involved in hydrolysis. These residues could aid in the development of inhibitors against this enzyme, thus developing novel antibiotics against Mycobacterium tuberculosis.

Publication Date

8-16-2024

Document Type

Thesis

Student Type

Graduate

Degree Name

Chemistry (MS)

Department, Program, or Center

Chemistry and Materials Science, School of

College

College of Science

Advisor

Suzanne O'Handley

Advisor/Committee Member

Michael Gleghorn

Advisor/Committee Member

Emiliano Brini

Comments

This thesis has been embargoed. The full-text will be available on or around 8/27/2025.

Campus

RIT – Main Campus

Download

Available for download on Wednesday, August 27, 2025

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