Subunit dimers of alpha-hemolysin expand the engineering toolbox for protein nanopores.

ABSTRACT: Staphylococcal α-hemolysin (αHL) forms a heptameric pore that features a 14-stranded transmembrane β-barrel. We attempted to force the αHL pore to adopt novel stoichiometries by oligomerizing subunit dimers generated by in vitro transcription and translation of a tandem gene. However, in vitro transcription and translation also produced truncated proteins, monomers, that were preferentially incorporated into oligomers. These oligomers were shown to be functional heptamers by single-channel recording and had a similar mobility to wild-type heptamers in SDS-polyacrylamide gels. Purified full-length subunit dimers were then prepared by using His-tagged protein. Again, single-channel recording showed that oligomers made from these dimers are functional heptamers, implying that one or more subu

SUBMITTER: Hammerstein AF 

PROVIDER: S-EPMC3077633 | biostudies-literature | 2011 Apr

REPOSITORIES: biostudies-literature