The cholesterol-dependent cytolysin signature motif: a critical element in the allosteric pathway that couples membrane binding to pore assembly.
The cholesterol-dependent cytolysins (CDCs) constitute a family of pore-forming toxins that contribute to the pathogenesis of a large number of Gram-positive bacterial pathogens.The most highly conserved region in the primary structure of the CDCs is the signature undecapeptide sequence (ECTGLAWEWWR). The CDC pore forming mechanism is highly sensitive to changes in its structure, yet its contribution to the molecular mechanism of the CDCs has remained enigmatic. Using a combination of fluorescence spectroscopic methods we provide evidence that shows the undecapeptide motif of the archetype CDC, perfringolysin O (PFO), is a key structural element in the allosteric coupling of the cholesterol-mediated membrane binding in domain 4 (D4) to distal structural changes in domain 3 (D3) that are required for the formation of the oligomeric pore complex. Loss of the undecapeptide function prevents all measurable D3 structural transitions, the intermolecular interaction of membrane bound monomers and the assembly of the oligomeric pore complex. We further show that this pathway does not exist in intermedilysin (ILY), a CDC that exhibits a divergent undecapeptide and that has evolved to use human CD59 rather than cholesterol as its receptor. These studies show for the first time that the undecapeptide of the cholesterol-binding CDCs forms a critical element of the allosteric pathway that controls the assembly of the pore complex.
Department of Microbiology and Immunology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
Kelley J
KJ
Dowd
Allison J
AJ
Farrand
Rodney K
RK
Tweten
D000494:Allosteric Regulation; D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001427:Bacterial Toxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001430:Bacteriocins / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002462:Cell Membrane / Q000382:microbiology*; D002784:Cholesterol / Q000378:metabolism*; D003016:Clostridium perfringens / Q000472:pathogenicity; D003603:Cytotoxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism* / Q000382:microbiology; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D017434:Protein Structure, Tertiary
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D001430:Bacteriocins; D003603:Cytotoxins; D006460:Hemolysin Proteins; C403910:intermedilysin protein, Streptococcus intermedius; C020452:Clostridium perfringens theta-toxin; D002784:Cholesterol