Oral Presentation The 3rd Prato Conference on Pore Forming Proteins 2015

Imaging the assembly of perfringolysin O (#8)

Michael J. Senior 1 , Alejandro P. Heuck 2 , Robert J.C. Gilbert 3 , Mark I. Wallace 1
  1. Chemistry Research Laboratory, University of Oxford, Oxford, United Kingdom
  2. Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA, USA
  3. Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, United Kingdom

Cholesterol-dependent cytolysins such as perfringolysin O (PFO) are proteins that lyse cells by forming large pores in the target cell membrane; they contribute to infections ranging from food poisoning to pneumonia and listeriosis. PFO monomers bind the target membrane and then oligomerize to form pores of 20-50 subunits, via a pre-pore intermediate. Here we study the assembly of these pores using single-molecule fluorescence imaging in Droplet Interface Bilayers. We track the increase in brightness as labelled monomers assemble to form individual pores, and observe intermediate stoichiometries during the assembly process.