Structural basis for tuning activity and membrane specificity of bacterial cytolysins

DOI: 10.1038/s41467-020-19482-6

Authors: Nita R. Shah (Imperial College London) , Tomas B. Voisin (Imperial College London) , Edward S. Parsons (University College London) , Courtney M. Boyd (Imperial College London) , Bart W. Hoogenboom (University College London) , Doryen Bubeck (Imperial College London)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 11

State: Published (Approved)
Published: November 2020
Diamond Proposal Number(s): 18659

Abstract: Cholesterol-dependent cytolysins (CDCs) are pore-forming proteins that serve as major virulence factors for pathogenic bacteria. They target eukaryotic cells using different mechanisms, but all require the presence of cholesterol to pierce lipid bilayers. How CDCs use cholesterol to selectively lyse cells is essential for understanding virulence strategies of several pathogenic bacteria, and for repurposing CDCs to kill new cellular targets. Here we address that question by trapping an early state of pore formation for the CDC intermedilysin, bound to the human immune receptor CD59 in a nanodisc model membrane. Our cryo electron microscopy map reveals structural transitions required for oligomerization, which include the lateral movement of a key amphipathic helix. We demonstrate that the charge of this helix is crucial for tuning lytic activity of CDCs. Furthermore, we discover modifications that overcome the requirement of cholesterol for membrane rupture, which may facilitate engineering the target-cell specificity of pore-forming proteins.

Journal Keywords: cryoEM; pore-forming proteins; CDC

Diamond Keywords: Bacteria

Subject Areas: Biology and Bio-materials, Chemistry

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios I-Titan Krios I at Diamond , Krios III-Titan Krios III at Diamond , Krios V-Titan Krios V at Diamond

Added On: 01/03/2021 12:01

Documents:
s41467-020-19482-6.pdf

Discipline Tags:

Pathogens Health & Wellbeing Biochemistry Chemistry Structural biology Life Sciences & Biotech

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)