An increasing number of bacterial pathogens are resistant to antibiotics. And the most dangerous pathogens share a common feature: a double membrane that is difficult to penetrate. Even when antibiotic agents are able to break into this shell, the bacteria just pump them right out again. But a recently discovered compound called Darobactin manages to circumvent these protective measures and kill nearly all problem pathogens. Researchers have now been able to elucidate the mechanism of action in a project funded by the Swiss National Science Foundation (SNSF) under the National Research Programme “Antimicrobial Resistance” (NRP 72).
Like a broken key
In a study published in Nature, the researchers describe how a deceptive manoeuvre enables Darobactin to work: its shape mimics a special three-dimensional structure normally found only in proteins produced by bacteria as building blocks for their outer membrane. The structure is the “key” for inserting the proteins into the outer shell at specific locations. Darobactin is a copy of this key. However, it does not act to penetrate the bacteria, but merely blocks the keyhole from the outside – like locking a door and then breaking off the key. As a result, the transport route for the bacteria’s shell components is obstructed and they die.
Hard to detect by conventional means
Similar mechanisms are already known in microbiology and are used by other drugs. The targeted binding structures, or keyholes, are usually quite large – at least in microbiological terms. In contrast, the target inhibited by Darobactin is very small and cannot be detected by conventional methods. At the same time, Darobactin is larger than most drugs and cannot get through the entry ports of the bacteria.
“At the beginning, we were puzzled by this,” say Sebastian Hiller and Timm Maier of the Biozentrum of the University of Basel, the two principal authors of the study. Their teams immediately realised that Darobactin does not act inside the pathogens but rather on the surface. There it interferes with the function of a protein known as BamA, which plays a central role in the construction of the double-protective membrane. “But how exactly Darobactin interacts with BamA was completely unclear,” says Hiller. It was only by combining several methods that the researchers finally identified the mechanism of action.
Spotting the perfect weak point
They discovered that Darobactin attacks a true Achilles heel of the pathogens: it binds directly to the most important site of BamA, the so-called backbone atoms. Because these atoms hold the protein together and determine its shape, it is nearly impossible to change them – though changing them would be the usual way for the bacteria to fend off a new antibiotic. In fact, Darobactin retained its effectiveness against all pathogens, for which Hiller and his team conducted laboratory tests that simulate resistance. In other words, the pathogens failed to change the broken lock.
Targeted drug development
These findings are a decisive step towards medical application, says infection biologist Dirk Bumann, who also conducts research at the Biozentrum in Basel. As co-director of the National Centre of Competence in Research (NCCR) “AntiResist”, which like NRP 72 is funded by the SNSF, he closely follows activities in antibiotics research. “Identifying Darobactin’s mechanism of action is a major achievement,” he says, “because it will enable further targeted improvement of Darobactin and its development into an effective drug”. That gives a boost to the long-cherished hope of finding a new generation of antibiotics to fight many of today’s problem pathogens.
Hundeep Kaur, Roman P. Jakob, Jan K. Marzinek, Robert Green, Yu Imai, Jani Reddy Bolla, Elia Agustoni, Carol V. Robinson, Peter J. Bond, Kim Lewis, Timm Maier & Sebastian Hiller, The antibiotic darobactin mimics a β-strand to inhibit outer membrane insertase. Nature; published online 14 April 2021
Press release published by the Swiss National Science Foundation (SNSF).