Assembly of α-Hemolysin: a Proteinaceous Pore with Potential Applications in Materials Synthesis

Abstract

Abstractα-Hemolysin (αHL) is secreted by the bacteriumStaphylococcus aureusas a watersoluble polypeptide of 293 amino acid residues. When presented with lipid bilayers or the detergent deoxycholate (DOC), aHL assembles into hexameric cylindrical pores that each contain one channel ∼ 1 to 2 nm in Internal diameter. A long-term goal of this laboratory is to use wild-type or re-engineered αHL pores as components of nanoscale materials: for example, to confer novel permeability properties upon thin films. The implementation of this concept would be facilitated by a better understanding of the mechanism by which the pore assembles. Reviewed here are findings that have given us insight Into the assembly mechanism, including the results of recent mutagenesis experiments. A critical summary is given of knowledge about the conformation of the monomer In solution, the hexamerIc pore and two proposed intermediates in assembly (a membrane-bound monomer and an oligomeric pore precursor). Future directions are outlined Including the prospects of obtaining three-dimensional structural data on the αHL pore or its precursors, methods for obtaining better monolayer sheets and new experiments on the topography of the pore and its precursors. The role of membrane receptors in facilitating the assembly of αHL is also discussed. Finally, it is demonstrated that despite our rather rudimentary knowledge of the assembly process, the Information gained so far still allows the design of mutant (αHL polypeptides with useful properties. For example, αHL mutants whose pore-forming ability is activated by proteases have been made.