Abstract
Cholesterol is believed to serve as the common receptor for the cholesterol-dependent cytolysins (CDCs). One member of this toxin family, Streptococcus intermedius intermedilysin (ILY), exhibits a narrow spectrum of cellular specificity that is seemingly inconsistent with this premise. We show here that ILY, via its domain 4 structure, binds to the glycosyl-phosphatidylinositol–linked membrane protein human CD59 (huCD59). CD59 is an inhibitor of the membrane attack complex of human complement. ILY specifically binds to huCD59 via residues that are the binding site for the C8α and C9 complement proteins. These studies provide a new model for the mechanism of cellular recognition by a CDC.
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References
Boulnois, G.J. Pneumococcal proteins and the pathogenesis of disease caused by Streptococcus pneumoniae. J. Gen. Microbiol. 138, 249–259 (1992).
Bricker, A.L., Cywes, C., Ashbaugh, C.D. & Wessels, M.R. NAD+-glycohydrolase acts as an intracellular toxin to enhance the extracellular survival of group A streptococci. Mol. Microbiol. 44, 257–269 (2002).
Sierig, G., Cywes, C., Wessels, M.R. & Ashbaugh, C.D. Cytotoxic effects of streptolysin O and streptolysin S enhance the virulence of poorly encapsulated group A streptococci. Infect. Immun. 71, 446–455 (2003).
Portnoy, D., Jacks, P.S. & Hinrichs, D. The role of hemolysin for intracellular growth of Listeria monocytogenes. J. Exp. Med. 167, 1459–1471 (1988).
Awad, M.M., Bryant, A.E., Stevens, D.L. & Rood, J.I. Virulence studies on chromosomal α-toxin and τ-toxin mutants constructed by allelic exchange provide genetic evidence for the essential role of α-toxin in Clostridium perfringens-mediated gas gangrene. Mol. Microbiol. 15, 191–202 (1995).
Billington, S.J., Jost, B.H., Cuevas, W.A., Bright, K.R. & Songer, J.G. The Arcanobacterium (Actinomyces) pyogenes hemolysin, pyolysin, is a novel member of the thiol-activated cytolysin family. J. Bacteriol. 179, 6100–6106 (1997).
Nagamune, H. et al. Intermedilysin, a novel cytotoxin specific for human cells secreted by Streptococcus intermedius UNS46 isolated from a human liver abscess. Infect. Immun. 64, 3093–3100 (1996).
Nagamune, H. et al. Distribution of the intermedilysin gene among the anginosus group streptococci and correlation between intermedilysin production and deep-seated infection with Streptococcus intermedius. J. Clin. Microbiol. 38, 220–226 (2000).
Olofsson, A., Hebert, H. & Thelestam, M. The projection structure of perfringolysin-O (Clostridium perfringens τ-toxin). FEBS Lett. 319, 125–127 (1993).
Shatursky, O. et al. The mechanism of membrane insertion for a cholesterol-dependent cytolysin: a novel paradigm for pore-forming toxins. Cell 99, 293–299 (1999).
Shepard, L.A. et al. Identification of a membrane-spanning domain of the thiol-activated pore-forming toxin Clostridium perfringens perfringolysin O: an α-helical to β-sheet transition identified by fluorescence spectroscopy. Biochemistry 37, 14563–14574 (1998).
Alouf, J.E. Introduction to the family of the structurally related cholesterol-binding cytolysins ('sulfhydryl-activated toxins'). In Bacterial Toxins: A Comprehensive Sourcebook (eds. Alouf, J. & Freer, J.) 443–456 (Academic Press, London, 1999).
Ohno-Iwashita, Y., Iwamoto, M., Ando, S., Mitsui, K. & Iwashita, S. A modified θ-toxin produced by limited proteolysis and methylation: a probe for the functional study of membrane cholesterol. Biochim. Biophys. Acta 1023, 441–448 (1990).
Geoffroy, C. & Alouf, J.E. Interaction of alveolysin A sulfhydryl-activated bacterial cytolytic toxin with thiol group reagents and cholesterol. Toxicon 20, 239–241 (1982).
Prigent, D. & Alouf, J.E. Interaction of streptolysin O with sterols. Biochem. Biophys. Acta. 433, 422–428 (1976).
Jacobs, T. et al. Listeriolysin O: cholesterol inhibits cytolysis but not binding to cellular membranes. Mol. Microbiol. 28, 1081–1089 (1998).
Giddings, K.S., Johnson, A.E. & Tweten, R.K. Redefining cholesterol's role in the mechanism of the cholesterol-dependent cytolysins. Proc. Natl. Acad. Sci. USA 100, 11315–11320 (2003).
Rollins, S.A. & Sims, P.J. The complement-inhibitory activity of CD59 resides in its capacity to block incorporation of C9 into membrane C5b-9. J. Immunol. 144, 3478–3483 (1990).
Rollins, S.A., Zhao, J., Ninomiya, H. & Sims, P.J. Inhibition of homologous complement by CD59 is mediated by a species-selective recognition conferred through binding to C8 within C5b-8 or C9 within C5b-9. J. Immunol. 146, 2345–2351 (1991).
Chang, C.P., Husler, T., Zhao, J., Wiedmer, T. & Sims, P.J. Identity of a peptide domain of human C9 that is bound by the cell-surface complement inhibitor, CD59. J. Biol. Chem. 269, 26424–26430 (1994).
Lockert, D.H. et al. Identity of the segment of human complement C8 recognized by complement regulatory protein CD59. J. Biol. Chem. 270, 19723–19728 (1995).
Gordon, V.M. et al. Clostridium septicum α-toxin uses glycosylphosphatidylinositol-anchored protein receptors. J. Biol. Chem. 274, 27274–27280 (1999).
Rudd, P.M. et al. The glycosylation of the complement regulatory protein, human erythrocyte CD59. J. Biol. Chem. 272, 7229–7244 (1997).
Petranka, J. et al. Structure-function relationships of the complement regulatory protein, CD59. Blood Cells Mol. Dis. 22, 281–296 (1996).
Ninomiya, H. et al. Contribution of the N-linked carbohydrate of erythrocyte antigen CD59 to its complement-inhibitory activity. J. Biol. Chem. 267, 8404–8410 (1992).
Ninomiya, H. & Sims, P.J. The human complement regulatory protein CD59 binds to the α-chain of C8 and to the 'b' domain of C9. J. Biol. Chem. 267, 13675–13680 (1992).
Zhang, H.F. et al. Identification of the individual residues that determine human CD59 species selective activity. J. Biol. Chem. 274, 10969–10974 (1999).
Zhao, X.J., Zhao, J., Zhou, Q. & Sims, P.J. Identity of the residues responsible for the species-restricted complement inhibitory function of human CD59. J. Biol. Chem. 273, 10665–10671 (1998).
Shimada, Y., Maruya, M., Iwashita, S. & Ohno-Iwashita, Y. The C-terminal domain of perfringolysin O is an essential cholesterol-binding unit targeting to cholesterol-rich microdomains. Eur. J. Biochem. 269, 6195–6203 (2002).
Weis, S. & Palmer, M. Streptolysin O: the C-terminal, tryptophan-rich domain carries functional sites for both membrane binding and self-interaction but not for stable oligomerization. Biochim. Biophys. Acta 1510, 292–299 (2001).
Fleming, T.J., O'Huigin, C. & Malek, T.R. Characterization of two novel Ly-6 genes. Protein sequence and potential structural similarity to α-bungarotoxin and other neurotoxins. J. Immunol. 150, 5379–5390 (1993).
Fletcher, C.M., Harrison, R.A., Lachmann, P.J. & Neuhaus, D. Structure of a soluble, glycosylated form of the human complement regulatory protein CD59. Structure 2, 185–199 (1994).
Nagamune, H. et al. Intermedilysin. A cytolytic toxin specific for human cells of a Streptococcus intermedius isolated from human liver abscess. Adv. Exp. Med. Biol. 418, 773–775 (1997).
Whiley, R.A., Beighton, D., Winstanley, T.G., Fraser, H.Y. & Hardie, J.M. Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus (the Streptococcus milleri group): association with different body sites and clinical infections. J. Clin. Microbiol. 30, 243–244 (1992).
Macey, M.G., Whiley, R.A., Miller, L. & Nagamune, H. Effect on polymorphonuclear cell function of a human-specific cytotoxin, intermedilysin, expressed by Streptococcus intermedius. Infect. Immun. 69, 6102–6109 (2001).
Gordon, D.L., Papazaharoudakis, H., Sadlon, T.A., Arellano, A. & Okada, N. Upregulation of human neutrophil CD59, a regulator of the membrane attack complex of complement, following cell activation. Immunol. Cell Biol. 72, 222–229 (1994).
Rother, R.P., Zhao, J., Zhou, Q. & Sims, P.J. Elimination of potential sites of glycosylation fails to abrogate complement regulatory function of cell surface CD59. J. Biol. Chem. 271, 23842–23845 (1996).
Melton, J.A., Parker, M.W., Rossjohn, J., Buckley, J.T. & Tweten, R.K. The identification and structure of the membrane-spanning domain of the Clostridium septicum α-toxin. J. Biol. Chem. 279, 14315–14322 (2004).
Laemmli, U.K. Cleavage of structural proteins during he assembly of the head of bacteriophage T4. Nature 227, 680–685 (1970).
Hotze, E.M. et al. Arresting pore formation of a cholesterol-dependent cytolysin by disulfide trapping synchronizes the insertion of the transmembrane β-sheet from a prepore intermediate. J. Biol. Chem. 276, 8261–8268 (2001).
Bodian, D.L., Davis, S.J., Morgan, B.P. & Rushmere, N.Ks. Mutational analysis of the active site and antibody epitopes of the complement-inhibitory glycoprotein, CD59. J. Exp. Med. 185, 507–516 (1997).
Acknowledgements
This work was supported by grants from the US National Institute of Allergy and Infectious Diseases (AI037657 and T32 AI07364), and from the US National Heart, Lung and Blood Institute (HL36061) The technical assistance of A. Marpoe and L. Bentsen was appreciated.
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Giddings, K., Zhao, J., Sims, P. et al. Human CD59 is a receptor for the cholesterol-dependent cytolysin intermedilysin. Nat Struct Mol Biol 11, 1173–1178 (2004). https://doi.org/10.1038/nsmb862
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DOI: https://doi.org/10.1038/nsmb862
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