title: An intermolecular electrostatic interaction controls the prepore-to-pore transition in a cholesterol-dependent cytolysin.
abstract: β-Barrel pore-forming toxins (βPFTs) form an obligatory oligomeric prepore intermediate before the formation of the β-barrel pore. The molecular components that control the critical prepore-to-pore transition remain unknown for βPFTs. Using the archetype βPFT perfringolysin O, we show that E183 of each monomer within the prepore complex forms an intermolecular electrostatic interaction with K336 of the adjacent monomer on completion of the prepore complex. The signal generated throughout the prepore complex by this interaction irrevocably commits it to the formation of the membrane-inserted giant β-barrel pore. This interaction supplies the free energy to overcome the energy barrier (determined here to be ∼ 19 kcal/mol) to the prepore-to-pore transition by the coordinated disruption of a critical interface within each monomer. These studies provide the first insight to our knowledge into the molecular mechanism that controls the prepore-to-pore transition for a βPFT.
authors: Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104;
authors: Kristin R
authors: KR
authors: Wade
authors: Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104;
authors: Eileen M
authors: EM
authors: Hotze
authors: Victorian Life Sciences Computation Initiative, The University of Melbourne, Parkville 3010, Victoria, Australia;
authors: Michael J
authors: MJ
authors: Kuiper
authors: Australian Cancer Research Foundation Rational Drug Discovery Centre, St. Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia; and.
authors: Craig J
authors: CJ
authors: Morton
authors: Australian Cancer Research Foundation Rational Drug Discovery Centre, St. Vincent's Institute of Medical Research, Fitzroy, Victoria 3065, Australia; and Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors: Michael W
authors: MW
authors: Parker
authors: Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104; Rod-Tweten@ouhsc.edu.
authors: Rodney K
authors: RK
authors: Tweten
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D056004:Molecular Dynamics Simulation; D009154:Mutation; D013050:Spectrometry, Fluorescence; D055672:Static Electricity; D013301:Streptolysins / Q000378:metabolism*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D013301:Streptolysins; C025964:streptolysin O; D002784:Cholesterol
keywords: alpha-hemolysin; hemolysin; sterol; streptolysin; toxin
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