Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is one of the deadliest infectious diseases globally. Antibiotics are available for TB however require a lengthy treatment timeframe of 4-6 months. (1) This is in-part due to the bacteria’s ability to switch to a dormant, non-replicating, phenotype which is inherently resistant to many antibiotics (2). Poor adherence to the lengthy treatment schedule has contributed to the consistent incidence of multiply-drug-resistant (MDR) and extensively-drug-resistant (XDR) TB cases over the past decade. (1,3) As such, there is an urgent need for new antibiotics which shorten treatment duration and operate via novel modes of action.(3)
Cyclomarin A, Ilamycin E and Ecumicin are cyclic peptide natural products and are some of the most potent antimycobacterial compounds to be discovered in recent years.(4) An analogue of Ecumicin, PH990 developed in house exhibited potent bactericidal activity against Mtb, capable of sterilizing cultures after seven days, and all three compounds, PH990, Cyclomarin A and Ilamycin E retain bactericidal activity against non-replicating Mtb.(5-7) These compounds bind to the N-terminal domain of ATPase chaperone ClpC1 and modulate ClpP1P2 facilitated protein degradation.(8) This presentation will describe the proteomics-based strategies which revealed the effects of the modulators on the entire proteome of Mtb. The compounds alter substrate recognition, simultaneously promoting the excessive degradation and accumulation of hundreds of different proteins. Despite structural similarities and overlapping binding sites, these compounds have strikingly different impacts on the abundance of specific proteins, including ClpC2 and Hsp.