# Dissection of Mycobacterium tuberculosis Clp protease assembly, activity and regulation

> **NIH NIH P20** · UNIVERSITY OF DELAWARE · 2020 · $232,501

## Abstract

Project Summary/Abstract
Drug-resistant tuberculosis is a global public health crisis. Mycobacterium tuberculosis (Mtb) causes ~1.3 million
deaths each year. 5% of new Mtb infections are drug-resistant, driving an urgent clinical need for new anti-Mtb
therapeutics. This proposal supports efforts to combat drug-resistant Mtb by dissecting the function and
pharmacological dysregulation of Clp proteases, a family of novel antibacterial targets. Clp proteases harness
chemical energy to destroy folded proteins in the cytosol of bacteria. Clp proteases are essential in mycobacteria,
and compounds that disrupt their function can kill Mtb and potentially treat drug-resistant tuberculosis. Our prior
work on Mtb Clp proteases revealed distinctive characteristics that set them apart from well-studied homologs.
We hypothesize that mycobacteria exploit these characteristics to regulate Clp protease assembly and activity
across stages of mycobacterial growth and infection. However, no studies have yet examined how these features
contribute to mycobacterial physiology, and this deficiency in our understanding hampers efforts to develop Clp-
targeting therapeutics. This sub-project aims to addresses outstanding questions surrounding Mtb Clp protease
regulation, and to develop compounds optimized to target Mtb Clp proteases. In Aim 1, we will interrogate Clp
protease regulation by investigating how and when Clp proteases form proteolytically active complexes. We will
use biophysical approaches to define the kinetic trajectory of assembly, and identify active and inactive complex
intermediates that form along the assembly pathway. Additionally, we will use a model mycobacterial system to
correlate assembly state and activity state within the cell. Understanding these details will improve our ability to
effectively target these enzymes during infection. In Aim 2, we seek to elaborate existing chemical scaffolds to
create new compounds optimized to target Mtb Clp enzymes. We will develop novel probes that modulate or
report on Clp protease assembly, and new lead compounds with improved affinity and target selectivity.
Structural information, together with our existing toolbox of biochemical assays, will guide compound
development and will be used to evaluate compound affinity and selectivity. We will also create and screen a
compound library, derived from an existing scaffold, to identify novel leads. Together, these studies will expand
our understanding of Mtb Clp protease function, and support the development of therapeutics capable of
combating drug-resistant tuberculosis.

## Key facts

- **NIH application ID:** 10026272
- **Project number:** 2P20GM104316-06A1
- **Recipient organization:** UNIVERSITY OF DELAWARE
- **Principal Investigator:** Karl Robert Schmitz
- **Activity code:** P20 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $232,501
- **Award type:** 2
- **Project period:** 2014-09-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10026272

## Citation

> US National Institutes of Health, RePORTER application 10026272, Dissection of Mycobacterium tuberculosis Clp protease assembly, activity and regulation (2P20GM104316-06A1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10026272. Licensed CC0.

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