Cholesterol Exposure at the Membrane Surface is Necessary and Sufficient to Trigger Perfringolysin O Binding

Overview

Journal Biochemistry

Specialty Biochemistry

Date 2009 Mar 19

PMID 19292457

Citations 73

Authors

John J Flanagan

Rodney K Tweten

Arthur E Johnson

Alejandro P Heuck

Affiliations

Soon will be listed here.

Abstract

Perfringolysin O (PFO) is the prototype for the cholesterol-dependent cytolysins, a family of bacterial pore-forming toxins that act on eukaryotic membranes. The pore-forming mechanism of PFO exhibits an absolute requirement for membrane cholesterol, but the complex interplay between the structural arrangement of the PFO C-terminal domain and the distribution of cholesterol in the target membrane is poorly understood. Herein we show that PFO binding to the bilayer and the initiation of the sequence of events that culminate in the formation of a transmembrane pore depend on the availability of free cholesterol at the membrane surface, while changes in the acyl chain packing of the phospholipids and cholesterol in the membrane core, or the presence or absence of detergent-resistant domains do not correlate with PFO binding. Moreover, PFO association with the membrane was inhibited by the addition of sphingomyelin, a typical component of membrane rafts in cell membranes. Finally, addition of molecules that do not interact with PFO, but intercalate into the membrane and displace cholesterol from its association with phospholipids (e.g., epicholesterol), reduced the amount of cholesterol required to trigger PFO binding. Taken together, our studies reveal that PFO binding to membranes is triggered when the concentration of cholesterol exceeds the association capacity of the phospholipids, and this cholesterol excess is then free to associate with the toxin.

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References

Soltani C, Hotze E, Johnson A, Tweten R . Specific protein-membrane contacts are required for prepore and pore assembly by a cholesterol-dependent cytolysin. J Biol Chem. 2007; 282(21):15709-16. PMC: 3746338. DOI: 10.1074/jbc.M701173200. View

Ramachandran R, Heuck A, Tweten R, Johnson A . Structural insights into the membrane-anchoring mechanism of a cholesterol-dependent cytolysin. Nat Struct Biol. 2002; 9(11):823-7. DOI: 10.1038/nsb855. View

London E, Brown D . Insolubility of lipids in triton X-100: physical origin and relationship to sphingolipid/cholesterol membrane domains (rafts). Biochim Biophys Acta. 2000; 1508(1-2):182-95. DOI: 10.1016/s0304-4157(00)00007-1. View

Nelson L, Johnson A, London E . How interaction of perfringolysin O with membranes is controlled by sterol structure, lipid structure, and physiological low pH: insights into the origin of perfringolysin O-lipid raft interaction. J Biol Chem. 2007; 283(8):4632-42. DOI: 10.1074/jbc.M709483200. View

Tilley S, Orlova E, Gilbert R, Andrew P, Saibil H . Structural basis of pore formation by the bacterial toxin pneumolysin. Cell. 2005; 121(2):247-56. DOI: 10.1016/j.cell.2005.02.033. View

Lange Y, Ye J, Steck T . Activation of membrane cholesterol by displacement from phospholipids. J Biol Chem. 2005; 280(43):36126-31. DOI: 10.1074/jbc.M507149200. View

Rosado C, Buckle A, Law R, Butcher R, Kan W, Bird C . A common fold mediates vertebrate defense and bacterial attack. Science. 2007; 317(5844):1548-51. DOI: 10.1126/science.1144706. View

ALOUF J, Geoffroy C, Pattus F, Verger R . Surface properties of bacterial sulfhydryl-activated cytolytic toxins. Interaction with monomolecular films of phosphatidylcholine and various sterols. Eur J Biochem. 1984; 141(1):205-10. DOI: 10.1111/j.1432-1033.1984.tb08176.x. View

Czajkowsky D, Hotze E, Shao Z, Tweten R . Vertical collapse of a cytolysin prepore moves its transmembrane beta-hairpins to the membrane. EMBO J. 2004; 23(16):3206-15. PMC: 514522. DOI: 10.1038/sj.emboj.7600350. View

10.

Alving C, Habig W, Urban K, HARDEGREE M . Cholesterol-dependent tetanolysin damage to liposomes. Biochim Biophys Acta. 1979; 551(1):224-8. DOI: 10.1016/0005-2736(79)90368-7. View

11.

Shepard L, Shatursky O, Johnson A, Tweten R . The mechanism of pore assembly for a cholesterol-dependent cytolysin: formation of a large prepore complex precedes the insertion of the transmembrane beta-hairpins. Biochemistry. 2000; 39(33):10284-93. DOI: 10.1021/bi000436r. View

12.

Andrich M, Vanderkooi J . Temperature dependence of 1,6-diphenyl-1,3,5-hexatriene fluorescence in phophoslipid artificial membranes. Biochemistry. 1976; 15(6):1257-61. DOI: 10.1021/bi00651a013. View

13.

Hamman B, Hendershot L, Johnson A . BiP maintains the permeability barrier of the ER membrane by sealing the lumenal end of the translocon pore before and early in translocation. Cell. 1998; 92(6):747-58. DOI: 10.1016/s0092-8674(00)81403-8. View

14.

Heuck A, Hotze E, Tweten R, Johnson A . Mechanism of membrane insertion of a multimeric beta-barrel protein: perfringolysin O creates a pore using ordered and coupled conformational changes. Mol Cell. 2000; 6(5):1233-42. DOI: 10.1016/s1097-2765(00)00119-2. View

15.

Heuck A, Savva C, Holzenburg A, Johnson A . Conformational changes that effect oligomerization and initiate pore formation are triggered throughout perfringolysin O upon binding to cholesterol. J Biol Chem. 2007; 282(31):22629-37. DOI: 10.1074/jbc.M703207200. View

16.

Rosenqvist E, Michaelsen T, Vistnes A . Effect of streptolysin O and digitonin on egg lecithin/cholesterol vesicles. Biochim Biophys Acta. 1980; 600(1):91-102. DOI: 10.1016/0005-2736(80)90414-9. View

17.

Soltani C, Hotze E, Johnson A, Tweten R . Structural elements of the cholesterol-dependent cytolysins that are responsible for their cholesterol-sensitive membrane interactions. Proc Natl Acad Sci U S A. 2007; 104(51):20226-31. PMC: 2154413. DOI: 10.1073/pnas.0708104105. View

18.

Polekhina G, Giddings K, Tweten R, Parker M . Insights into the action of the superfamily of cholesterol-dependent cytolysins from studies of intermedilysin. Proc Natl Acad Sci U S A. 2005; 102(3):600-5. PMC: 545513. DOI: 10.1073/pnas.0403229101. View

19.

Hadders M, Beringer D, Gros P . Structure of C8alpha-MACPF reveals mechanism of membrane attack in complement immune defense. Science. 2007; 317(5844):1552-4. DOI: 10.1126/science.1147103. View

20.

Ohno-Iwashita Y, Iwamoto M, Ando S, Iwashita S . Effect of lipidic factors on membrane cholesterol topology--mode of binding of theta-toxin to cholesterol in liposomes. Biochim Biophys Acta. 1992; 1109(1):81-90. DOI: 10.1016/0005-2736(92)90190-w. View