3 research outputs found
Combined Structural Analysis and Molecular Dynamics Reveal Penicillin-Binding Protein Inhibition Mode with β‑Lactones
β-Lactam antibiotics comprise one of the most widely
used
therapeutic classes to combat bacterial infections. This general scaffold
has long been known to inhibit bacterial cell wall biosynthesis by
inactivating penicillin-binding proteins (PBPs); however, bacterial
resistance to β-lactams is now widespread, and new strategies
are urgently needed to target PBPs and other proteins involved in
bacterial cell wall formation. A key requirement in the identification
of strategies to overcome resistance is a deeper understanding of
the roles of the PBPs and their associated proteins during cell growth
and division, such as can be obtained with the use of selective chemical
probes. Probe development has typically depended upon known PBP inhibitors,
which have historically been thought to require a negatively charged
moiety that mimics the C-terminus of the PBP natural peptidoglycan
substrate, d-Ala-d-Ala. However, we have identified
a new class of β-lactone-containing molecules that interact
with PBPs, often in an isoform-specific manner, and do not incorporate
this C-terminal mimetic. Here, we report a series of structural biology
experiments and molecular dynamics simulations that we utilized to
evaluate specific binding modes of this novel PBP inhibitor class.
In this work, we obtained <2 Å resolution X-ray structures
of four β-lactone probes bound to PBP1b from Streptococcus
pneumoniae. Despite their diverging recognition modes beyond
the site of covalent modification, these four probes all efficiently
labeled PBP1b, as well as other PBPs from S. pneumoniae. From these structures, we analyzed protein–ligand
interactions and characterized the β-lactone-bound active sites
using in silico mutagenesis and molecular dynamics.
Our approach has clarified the dynamic interaction profile in this
series of ligands, expanding the understanding of PBP inhibitor binding
Dual Inhibitor of MurD and MurE Ligases from <i>Escherichia coli</i> and <i>Staphylococcus aureus</i>
MurD and MurE ligases, consecutive enzymes participating
in the
intracellular steps of bacterial peptidoglycan biosynthesis, are important
targets for antibacterial drug discovery. We have designed, synthesized,
and evaluated the first d-glutamic acid-containing dual inhibitor
of MurD and MurE ligases from <i>Escherichia coli</i> and <i>Staphylococcus aureus</i> (IC<sub>50</sub> values between 6.4
and 180 μM) possessing antibacterial activity against Gram-positive <i>S. aureus</i> and its methicillin-resistant strain (MRSA) with
minimal inhibitory concentration (MIC) values of 8 μg/mL. The
inhibitor was also found to be noncytotoxic for human HepG2 cells
at concentrations below 200 μM