title: Field-dependent effect of crown ether (18-crown-6) on ionic conductance of alpha-hemolysin channels.
abstract: Closing linear poly(ethylene glycol) (PEG) into a circular "crown" dramatically changes its dynamics in the alpha-hemolysin channel. In the electrically neutral crown ether (C2H4O)6, six ethylene oxide monomers are linked into a circle that gives the molecule ion-complexing capacity and increases its rigidity. As with linear PEG, addition of the crown to the membrane-bathing solution decreases the ionic conductance of the channel and generates additional conductance noise. However, in contrast to linear PEG, both the conductance reduction (reporting on crown partitioning into the channel pore) and the noise (reporting on crown dynamics in the pore) now depend on voltage strongly and nonmonotonically. Within the whole frequency range accessible in channel reconstitution experiments, the noise power spectrum is "white", showing that crown exchange between the channel and the bulk solution is fast. Analyzing these data in the framework of a Markovian two-state model, we are able to characterize the process quantitatively. We show that the lifetime of the crown in the channel reaches its maximum (a few microseconds) at about the same voltage (approximately 100 mV, negative from the side of protein addition) where the crown's reduction of the channel conductance is most pronounced. Our interpretation is that, because of its rigidity, the crown feels an effective steric barrier in the narrowest part of the channel pore. This barrier together with crown-ion complexing and resultant interaction with the applied field leads to behavior usually associated with voltage-dependent binding in the channel pore.
authors.affiliation: Laboratory of Physical and Structural Biology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Sergey M
authors.initials: SM
authors.lastname: Bezrukov
authors.forename: Oleg V
authors.initials: OV
authors.lastname: Krasilnikov
authors.forename: Liliya N
authors.initials: LN
authors.lastname: Yuldasheva
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Berezhkovskii
authors.forename: Claudio G
authors.initials: CG
authors.lastname: Rodrigues
mesh_terms: D003198:Computer Simulation; D043844:Crown Ethers / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity; D004574:Electromagnetic Fields; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000187:drug effects / Q000528:radiation effects; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000187:drug effects / Q000528:radiation effects; D015640:Ion Channel Gating / Q000187:drug effects* / Q000528:radiation effects*; D007477:Ions; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry* / Q000528:radiation effects; D008564:Membrane Potentials / Q000187:drug effects / Q000528:radiation effects; D008567:Membranes, Artificial; D008956:Models, Chemical; D008958:Models, Molecular
chemical_list: D043844:Crown Ethers; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D007477:Ions; D008051:Lipid Bilayers; D008567:Membranes, Artificial; C015762:18-crown-6