Monomer-monomer interactions propagate structural transitions necessary for pore formation by the cholesterol-dependent cytolysins

J Biol Chem. 2012 Jul 13;287(29):24534-43. doi: 10.1074/jbc.M112.380139. Epub 2012 May 29.

Abstract

The assembly of the cholesterol-dependent cytolysin (CDC) oligomeric pore complex requires a complex choreography of secondary and tertiary structural changes in domain 3 (D3) of the CDC monomer structure. A point mutation was identified in the archetype CDC, perfringolysin O, that blocks detectable D3 structural changes and traps the membrane-bound monomers in an early and reversible stage of oligomer assembly. Using this and other mutants we show that specific D3 structural changes are propagated from one membrane-bound monomer to another. Propagation of these structural changes results in the exposure of a β-strand in D3 that allows it to pair and form edge-on interactions with a second β-strand of a free membrane-bound monomer. Pairing of these strands establishes the final geometry of the pore complex and is necessary to drive the formation of the β-barrel pore. These studies provide new insights into how structural information is propagated between membrane-bound monomers of a self-assembling system and the interactions that establish the geometry of the final pore complex.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Cholesterol / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescence Resonance Energy Transfer
  • Hemolysis
  • Humans
  • Microscopy, Electron
  • Perforin / chemistry*
  • Perforin / genetics
  • Perforin / metabolism*
  • Point Mutation / genetics
  • Protein Structure, Secondary
  • Protein Structure, Tertiary

Substances

  • Perforin
  • Cholesterol