Details

Autor(en) / Beteiligte

• Goult, Jonathan D
• Van, Daniel C L
• Taylor, Yasmin V
• Inns, Patrick G
• Kaminska, Renata
• Vesely, Martin
• Kleanthous, Colin
• Paci, Emanuele

Titel

Structural constraints of pyocin S2 import through the ferripyoverdine receptor FpvAI

Ist Teil von

• PNAS nexus, 2024-04, Vol.3 (4), p.pgae124-pgae124

Ort / Verlag

England: Oxford University Press

Erscheinungsjahr

2024

Link zum Volltext

Quelle

Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals

Beschreibungen/Notizen

• TonB-dependent transporters (TBDTs) mediate energized transport of essential nutrients into gram-negative bacteria. TBDTs are increasingly being exploited for the delivery of antibiotics to drug-resistant bacteria. While much is known about ground state complexes of TBDTs, few details have emerged about the transport process itself. In this study, we exploit bacteriocin parasitization of a TBDT to probe the mechanics of transport. Previous work has shown that the N-terminal domain of -specific bacteriocin pyocin S2 (PyoS2 ) is imported through the pyoverdine receptor FpvAI. PyoS2 transport follows the opening of a proton-motive force-dependent pore through FpvAI and the delivery of its own TonB box that engages TonB. We use molecular models and simulations to formulate a complete translocation pathway for PyoS2 that we validate using protein engineering and cytotoxicity measurements. We show that following partial removal of the FpvAI plug domain which occludes the channel, the pyocin's N-terminus enters the channel by electrostatic steering and ratchets to the periplasm. Application of force, mimicking that exerted by TonB, leads to unraveling of PyoS2 as it squeezes through the channel. Remarkably, while some parts of PyoS2 must unfold, complete unfolding is not required for transport, a result we confirmed by disulfide bond engineering. Moreover, the section of the FpvAI plug that remains embedded in the channel appears to serve as a buttress against which PyoS2 is pushed to destabilize the domain. Our study reveals the limits of structural deformation that accompanies import through a TBDT and the role the TBDT itself plays in accommodating transport.