Structural basis for tuning activity and membrane specificity of bacterial cytolysins

Nature Communications
Nita R ShahDoryen Bubeck

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.

References

Mar 14, 1997·The Journal of Biological Chemistry·P M RuddR A Dwek
Aug 4, 2001·The Journal of Biological Chemistry·D M HooverJ Lubkowski
Oct 2, 2003·The Journal of Biological Chemistry·Deborah A FraserB Paul Morgan
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Nov 16, 2004·Nature Structural & Molecular Biology·Kara S GiddingsRodney K Tweten
Jan 8, 2005·Proceedings of the National Academy of Sciences of the United States of America·Galina PolekhinaMichael W Parker
Sep 24, 2005·Infection and Immunity·Rodney K Tweten
Aug 29, 2006·Nature Reviews. Molecular Cell Biology·Robert B AbramovitchGregory B Martin
Mar 3, 2007·Journal of Molecular Biology·Jamie RossjohnMichael W Parker
Aug 3, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Kirstin J LeathSusan M Lea
Nov 9, 2007·Cellular and Molecular Life Sciences : CMLS·M R GonzalezB Frêche
Feb 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Allison J FarrandRodney K Tweten
Apr 13, 2010·Acta Crystallographica. Section D, Biological Crystallography·P EmsleyK Cowtan
Sep 8, 2010·Protein Engineering, Design & Selection : PEDS·Yelena V GrinkovaStephen G Sligar
Jan 5, 2011·Protein & Cell·Lingfeng XuZihe Rao
Apr 5, 2011·Acta Crystallographica. Section D, Biological Crystallography·Garib N MurshudovAlexei A Vagin
Nov 26, 2014·Proceedings of the National Academy of Sciences of the United States of America·Lucy K ShewellMichael P Jennings
Feb 6, 2015·PLoS Biology·Natalya LukoyanovaMichelle A Dunstone
Aug 19, 2015·Journal of Structural Biology·Alexis Rohou, Nikolaus Grigorieff
Dec 8, 2015·Nature Reviews. Microbiology·Matteo Dal Peraro, F Gisou van der Goot
Dec 3, 2016·Scientific Reports·Courtney M BoydDoryen Bubeck
Apr 27, 2018·Nature·Jianbin RuanHao Wu
Jun 7, 2018·Acta Crystallographica. Section D, Structural Biology·Pavel V AfoninePaul D Adams
Sep 11, 2018·Acta Crystallographica. Section D, Structural Biology·Pavel V AfonineAlexandre Urzhumtsev
Jun 19, 2019·Proceedings of the National Academy of Sciences of the United States of America·Martin VögeleGerhard Hummer
Jun 27, 2019·Communications Biology·Thorsten WagnerStefan Raunser
Sep 21, 2019·Nature Communications·Siew Siew PangJames C Whisstock
Nov 2, 2019·Nature·Shabih ShakeelLori A Passmore
Nov 7, 2019·Nature Reviews. Immunology·Petr BrozFeng Shao
Nov 30, 2019·Nature Communications·Jesse A Rudd-SchmidtIlia Voskoboinik

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Citations

Oct 1, 2021·Microbiology and Immunology·Atsushi Tabata, Hideaki Nagamune
Nov 9, 2021·Biochemical Society Transactions·Joshua T Benton, Charles Bayly-Jones

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Methods Mentioned

BETA
electron microscopy
atomic force microscopy
AFM
size exclusion chromatography

Software Mentioned

Namdinator
Refmac
EPU
SPM
Phenix real _ space _ refine
Gwyddion
Phenix
COOT
CTFFIND4
MotionCor2

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