Caseinolytic protease (ClpP) is a cylindrical protease that degrades proteins in the presence of ATPase chaperones. On its own, bacterial ClpP can only degrade small peptides; however, the addition of a novel class of antibiotics, ADEPs, can cause unregulated proteolysis leading to bacterial cell death. Bacterial ClpP is an attractive target for antibiotic development. A high-throughput screen of small molecules identified a group of compounds which are termed Activators of Self-Compartmentalizing Proteases (ACP). A collection of ACP3 and ACP4/5 analogs was synthesized and investigated for biological activity. The project resulted in compounds with greater activity than the lead structures against isolated E. coli ClpP. Also, several analogs possessed bacteriostatic activity against N. meningitidis and S. aureus cell lines.

Caseinolytic protease (ClpP) is a cylindrical protease that degrades proteins in the presence of ATPase chaperones. On its own, bacterial ClpP can only degrade small peptides; however, the addition of a novel class of antibiotics, ADEPs, can cause unregulated proteolysis leading to bacterial cell death. Bacterial ClpP is an attractive target for antibiotic development. A high-throughput screen of small molecules identified a group of compounds which are termed Activators of Self-Compartmentalizing Proteases (ACP). A collection of ACP3 and ACP4/5 analogs was synthesized and investigated for biological activity. The project resulted in compounds with greater activity than the lead structures against isolated E. coli ClpP. Also, several analogs possessed bacteriostatic activity against N. meningitidis and S. aureus cell lines.

Caseinolytic protease (ClpP) is a cylindrical protease forming the core of protein degradation machinery in eubacteria. ClpP is tightly regulated and is non-functional without a member of the Clp-ATPases. A new class of antibiotics, termed ADEPs, bind to ClpP and allow for activation without the Clp-ATPases; leading to cell death. A more efficient synthetic route to the ADEPs utilizing solid-phase peptide synthesis was investigated. A linear peptide was synthesized, however attempts to close the depsipeptidic macrocycle via macrolactonization failed. Further attempts of assembling a branched depsipeptide for ring closure via a macrolactamization resulted in products that were not stable to cleavage conditions. A group of molecules termed Activators of Self-Compartmentalizing Proteases (ACP) were identified through a screen for activity towards ClpP. Compound ACP1 was synthesized along with twelve analogs and their activity towards ClpP evaluated. The project resulted in a compound with a higher activity than its natural product counterpart.

Caseinolytic protease (ClpP) is a cylindrical protease forming the core of protein degradation machinery in eubacteria. ClpP is tightly regulated and is non-functional without a member of the Clp-ATPases. A new class of antibiotics, termed ADEPs, bind to ClpP and allow for activation without the Clp-ATPases; leading to cell death. A more efficient synthetic route to the ADEPs utilizing solid-phase peptide synthesis was investigated. A linear peptide was synthesized, however attempts to close the depsipeptidic macrocycle via macrolactonization failed. Further attempts of assembling a branched depsipeptide for ring closure via a macrolactamization resulted in products that were not stable to cleavage conditions. A group of molecules termed Activators of Self-Compartmentalizing Proteases (ACP) were identified through a screen for activity towards ClpP. Compound ACP1 was synthesized along with twelve analogs and their activity towards ClpP evaluated. The project resulted in a compound with a higher activity than its natural product counterpart.

Tuberculosis is a severe disease that causes about 2 million deaths every year. It is a worldwide threat and it is estimated that one-third of the world’s population carries the infection. The severe side effects of the present drugs, and the more than 6 months long treatment, in addition to the development of resistant bacterial strains, are the incentives for the intensified search for new drugs. In this work two potential mycobacterial drug targets have been studied: Caseinolytic protease 1 (ClpP1) from Mycobacterium tuberculosis (Mt) and Methionyl-tRNA synthetase (MetRS) from Mycobacterium smegmatis (Ms). The X-ray stucture of ClpP1 was determined to 3.0 Å resolution. The study gives details on the tetradecameric arrangement of the enzyme. Two hepameric discs assemble to form a chamber containing the catalytic activity mediated by each of the monomers. The chamber can be reached by two pores. Comparison with the human homologue reveals important structural differences. The X-ray studies on Ms MetRS were done to 2.3 Å and 2.8 Å resolution. The study gives details on the flexibility of the enzyme and how this is related to activity. Important findings are identification of an intermediate structure in which the methionine to be adenylated is bound in the catalytic site in a tight complex. The catalytic site and the anticodon recognizing domains are separated and the structural results indicate communication between the domains. The possibility to allosterically inhibit the enzyme is discussed.