Conformational cycle of a protease-containing ABC transporter in lipid nanodiscs reveals the mechanism of cargo-protein coupling

Protease-containing ABC transporters (PCATs) couple the energy of ATP hydrolysis to the processing and export of diverse cargo proteins across cell membranes to mediate antimicrobial resistance and quorum sensing. Here, we combine biochemical analysis, single particle cryoEM, and DEER spectroscopy i...

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Veröffentlicht in:Nature communications 2024-10, Vol.15 (1), p.9055-15, Article 9055
Hauptverfasser: Zhang, Ruojing, Jagessar, Kevin L., Brownd, Matthew, Polasa, Adithya, Stein, Richard A., Moradi, Mahmoud, Karakas, Erkan, Mchaourab, Hassane S.
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Sprache:eng
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Zusammenfassung:Protease-containing ABC transporters (PCATs) couple the energy of ATP hydrolysis to the processing and export of diverse cargo proteins across cell membranes to mediate antimicrobial resistance and quorum sensing. Here, we combine biochemical analysis, single particle cryoEM, and DEER spectroscopy in lipid bilayers along with computational analysis to illuminate the structural and energetic underpinnings of coupled cargo protein export. Our integrated investigation uncovers competitive interplay between nucleotides and cargo protein binding that ensures the latter’s orderly processing and subsequent transport. The energetics of cryoEM structures in lipid bilayers are congruent with the inferred mechanism from ATP turnover analysis and reveal a snapshot of a high-energy outward-facing conformation that provides an exit pathway into the lipid bilayer and/or the extracellular side. DEER investigation of the core ABC transporter suggests evolutionary tuning of the energetic landscape to fulfill the function of substrate processing prior to export. Protease containing ABC Transporters (PCAT) play a critical role in the translocation of polypeptides across membranes. Here, authors reveal the structural and energetic of the ATP-powered conformational cycle that enable this process.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-024-53420-0