Saturday, February 15, 2025

Protein shuttling mechanism helps bacteria pump out antibiotics and virulence factors

Amazing stuff! Are we cracking the defenses of bacteria? Is this the bacterial immune system?

"Just as bailing out water with a bucket can save a sinking boat, certain cells have their own special technology for expelling toxins. Unfortunately, “toxins” can include antibiotic treatments. ...

uncovered the equipment that enables bacteria to survive exposure to antibiotics: a shuttling mechanism that helps a complex of proteins pump out a wide spectrum of antibiotics along with other physiological substrates from the cell. ...

Some bacteria are more resistant to antibiotics than others, with so-called gram-negative bacteria being particularly resilient because they have an extra membrane to armor themselves. They also have a sophisticated plumbing system in the form of a three-part protein complex – MacAB-TolC – that spans the cell’s inner and outer membranes, as well as the periplasm that connects them. Each of the three key proteins occupies a different location: TolC on the outer membrane, MacB on the inner membrane and MacA in the periplasm, although it is anchored on the inner membrane.

This “tripartite” protein complex, also known as a multidrug efflux pump, forms a conduit that drains out not only antibiotics but also virulence factors – i.e., molecules that are produced by the bacterial cell itself and can infect or otherwise compromise its host. ..."

From the highlights and abstract:
"Highlights
• Single-molecule imaging reveals imbalanced stoichiometry of MacAB-TolC components in vivo
• Excess MacB shows spatiotemporal behaviors for efficient substrate sequestration and efflux
• The limiting MacA can disassemble from MacAB-TolC and shuttle among clustered MacB
• Chemical or physical perturbation can compromise MacAB-TolC function
Summary
Multidrug efflux pumps confer not only antibiotic resistance to bacteria but also cell proliferation.
In gram-negative bacteria, the ATP-binding cassette (ABC)-family transporter MacB, the adaptor protein MacA, and the outer membrane protein TolC form the MacA6:MacB2:TolC3 assembly to extrude antibiotics and virulence factors.
Here, using quantitative single-molecule single-cell imaging, we uncover that, in E. coli cells, there is a large excess of MacB (and TolC) driving the limiting adaptor protein MacA mostly into the MacAB-TolC assembly.
Moreover, the excess MacB transporters can dynamically cluster around the assembly, and MacA can dynamically disassemble from the MacAB-TolC assembly, leading to an adaptor protein shuttling mechanism for efficient substrate sequestration from the periplasm toward efflux.
We further show that both MacB clustering and MacAB-TolC assembly can be perturbed chemically or physically via microfluidics-based extrusion loading for compromised antibiotic tolerance. These insights may provide opportunities for countering the activities of multidrug efflux systems for antimicrobial treatments."

Protein shuttling mechanism helps bacteria pump out antibiotics | Cornell Chronicle



Graphical abstract


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