title: Structural elements of the cholesterol-dependent cytolysins that are responsible for their cholesterol-sensitive membrane interactions.
abstract: The pore-forming mechanism of the cholesterol-dependent cytolysins (CDCs) exhibits an absolute requirement for membrane cholesterol. The structural elements of the CDCs that mediate this interaction are not well understood. Three short hydrophobic loops (L1-L3) and a highly conserved undecapeptide sequence at the tip of domain 4 of the CDC structure are known to anchor the CDC to the membrane. It has been thought that the undecapeptide directly mediates the interaction of the CDCs with a cholesterol-rich cell surface. Herein we show that the L1-L3 loops, not the undecapeptide, are responsible for mediating the specific interaction of the CDCs with cholesterol-rich membranes. The membrane insertion of the undecapeptide was uncoupled from membrane binding by the covalent modification of the undecapeptide cysteine thiol. Modification of the cysteine prevented prepore to pore conversion, but did not affect membrane binding, thus demonstrating that undecapeptide membrane insertion follows that of the L1-L3 loops. These studies provide an example of a structural motif that specifically mediates the interaction of a bacterial toxin with a cholesterol-rich membrane.
authors: Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
authors: Casie E
authors: CE
authors: Soltani
authors: Eileen M
authors: EM
authors: Hotze
authors: Arthur E
authors: AE
authors: Johnson
authors: Rodney K
authors: RK
authors: Tweten
mesh_terms: D019943:Amino Acid Substitution; D001224:Aspartic Acid / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D001427:Bacterial Toxins / Q000737:chemistry* / Q000235:genetics; D001430:Bacteriocins / Q000737:chemistry* / Q000235:genetics; D002784:Cholesterol / Q000737:chemistry*; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry; D003603:Cytotoxins / Q000737:chemistry* / Q000235:genetics; D005998:Glycine / Q000737:chemistry; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000235:genetics; D008081:Liposomes / Q000737:chemistry; D008567:Membranes, Artificial; D016062:Porosity; D011487:Protein Conformation; D014364:Tryptophan / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D001430:Bacteriocins; D003603:Cytotoxins; D006460:Hemolysin Proteins; D008081:Liposomes; D008567:Membranes, Artificial; C403910:intermedilysin protein, Streptococcus intermedius; D001224:Aspartic Acid; C020452:Clostridium perfringens theta-toxin; D014364:Tryptophan; D002784:Cholesterol; D003545:Cysteine; D005998:Glycine