Exploiting the aggregation propensity of beta-lactamases to design inhibitors that induce enzyme misfolding

There is an arms race between beta-lactam antibiotics development and co-evolving beta-lactamases, which provide resistance by breaking down beta-lactam rings. We have observed that certain beta-lactamases tend to aggregate, which persists throughout their evolution under the selective pressure of antibiotics on their active sites. Interestingly, we find that existing beta-lactamase active site inhibitors can act as molecular chaperones, promoting the proper folding of these resistance factors. Therefore, we have created Pept-Ins, synthetic peptides designed to exploit the structural weaknesses of beta-lactamases by causing them to misfold into intracellular inclusion bodies. This approach restores sensitivity to a wide range of beta-lactam antibiotics in resistant clinical isolates, including those with Extended Spectrum variants that pose significant challenges in medical practice. Our findings suggest that targeted aggregation of resistance factors could offer a strategy for identifying molecules that aid in addressing the global antibiotic resistance crisis.

Ladan Khodaparast, Laleh Khodaparast, Guiqin Wu, Emiel Michiels, Rodrigo Gallardo, Bert Houben, Teresa Garcia, Matthias De Vleeschouwer, Meine Ramakers, Hannah Wilkinson, Ramon Duran-Romaña, Johan Van Eldere, Frederic Rousseau, Joost Schymkowitz