# Comparative structural and functional analysis of Pseudomonas aeruginosa inhibitor of vertebrate lysozyme paralogs

> **NIH NIH R16** · KENNESAW STATE UNIVERSITY · 2023 · $135,500

## Abstract

Project Summary – Comparative structural and functional analysis of Pseudomonas aeruginosa inhibitor
of vertebrate lysozyme paralogs.
Thomas Leeper, PI, Kennesaw State University.
The proposed integrative studies will investigate a significant regulator of peptidoglycan biochemistry in the
cystic fibrosis pathogen Pseudomonas aeruginosa (PA), known to display multidrug resistance
(MDR). Specifically, Inhibitor of Vertebrate Lysozyme protein 2 (IVYp2) will have its structure determined
by NMR (Aim 1) and compared to its ability to inhibit hydrolase enzyme kinetics (Aim 2). While IVY
proteins were originally suggested to inhibit lysozyme glycoside hydrolases, subsequent work suggested
that the evolutionary origin of IVY’s may have been to inhibit lytic transglycosylases (LTs). For example,
IVYp2 has no effect on lysozyme activity in enzyme inhibition assays but has been shown to inhibit LTs,
at least in vitro. In order to determine which LTs are recognized in vivo enzyme inhibition assays with
LTs (Aim 2) will be compared to novel applications of complementation and genetically encoded
crosslinking studies in IVY knockout PA strains (Aim 3). The identified cognate IVY-LT pairs will be
compared to the NMR structure of IVYp2 (Aim 1) and existing complex structures for other IVYs (i.e.
IVYp1) bound to lysozyme to suggest regions important for LT molecular recognition. Graduate and
undergraduate students, particularly those from underrepresented groups, will significantly participate in
this research to facilitate data collection while providing them important training and advancement
opportunities. These innovative studies will establish the background and workflow needed for future
drug discovery efforts to develop new drug classes targeting novel biochemical pathways in MDR PA.

## Key facts

- **NIH application ID:** 10691290
- **Project number:** 5R16AI167815-02
- **Recipient organization:** KENNESAW STATE UNIVERSITY
- **Principal Investigator:** THOMAS C LEEPER
- **Activity code:** R16 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $135,500
- **Award type:** 5
- **Project period:** 2022-08-26 → 2026-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10691290

## Citation

> US National Institutes of Health, RePORTER application 10691290, Comparative structural and functional analysis of Pseudomonas aeruginosa inhibitor of vertebrate lysozyme paralogs (5R16AI167815-02). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10691290. Licensed CC0.

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