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Single molecule FRET reveals pore size and opening mechanism of a mechano-sensitive ion channel.


ABSTRACT: The mechanosensitive channel of large conductance, which serves as a model system for mechanosensitive channels, has previously been crystallized in the closed form, but not in the open form. Ensemble measurements and electrophysiological sieving experiments show that the open-diameter of the channel pore is >25 Å, but the exact size and whether the conformational change follows a helix-tilt or barrel-stave model are unclear. Here we report measurements of the distance changes on liposome-reconstituted MscL transmembrane ?-helices, using a 'virtual sorting' single-molecule fluorescence energy transfer. We observed directly that the channel opens via the helix-tilt model and the open pore reaches 2.8 nm in diameter. In addition, based on the measurements, we developed a molecular dynamics model of the channel structure in the open state which confirms our direct observations. DOI: http://dx.doi.org/10.7554/eLife.01834.001.

SUBMITTER: Wang Y 

PROVIDER: S-EPMC3925968 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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Single molecule FRET reveals pore size and opening mechanism of a mechano-sensitive ion channel.

Wang Yong Y   Liu Yanxin Y   Deberg Hannah A HA   Nomura Takeshi T   Hoffman Melinda Tonks MT   Rohde Paul R PR   Schulten Klaus K   Martinac Boris B   Selvin Paul R PR  

eLife 20140218


The mechanosensitive channel of large conductance, which serves as a model system for mechanosensitive channels, has previously been crystallized in the closed form, but not in the open form. Ensemble measurements and electrophysiological sieving experiments show that the open-diameter of the channel pore is >25 Å, but the exact size and whether the conformational change follows a helix-tilt or barrel-stave model are unclear. Here we report measurements of the distance changes on liposome-recons  ...[more]

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