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    Imaging the Electrostatic Potential of Transmembrane Channels: Atomic Probe Microscopy of OmpF Porin

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    Issue Date
    2002-03
    Author
    Philippsen, Ansgar
    Engel, Andreas
    Schirmer, Tilman
    Roux, Benoît
    Müller, Daniel J.
    Im, Wonpil
    Publisher
    Elsevier
    Type
    Article
    Article Version
    Scholarly/refereed, publisher version
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    Abstract
    The atomic force microscope (AFM) was used to image native OmpF porin and to detect the electrostatic potential generated by the protein. To this end the OmpF porin trimers from Escherichia coli was reproducibly imaged at a lateral resolution of ∼0.5 nm and a vertical resolution of ∼0.1 nm at variable electrolyte concentrations of the buffer solution. At low electrolyte concentrations the charged AFM probe not only contoured structural details of the membrane protein surface but also interacted with local electrostatic potentials. Differences measured between topographs recorded at variable ionic strength allowed mapping of the electrostatic potential of OmpF porin. The potential map acquired by AFM showed qualitative agreement with continuum electrostatic calculations based on the atomic OmpF porin embedded in a lipid bilayer at the same electrolyte concentrations. Numerical simulations of the experimental conditions showed the measurements to be reproduced quantitatively when the AFM probe was included in the calculations. This method opens a novel avenue to determine the electrostatic potential of native protein surfaces at a lateral resolution better than 1 nm and a vertical resolution of ∼0.1 nm.
    Description
    This is the published version. Copyright 2002 by Elsevier.
    URI
    http://hdl.handle.net/1808/17440
    DOI
    https://doi.org/10.1016/S0006-3495(02)75517-3
    Collections
    • Molecular Biosciences Scholarly Works [581]
    Citation
    Philippsen, Ansgar, Wonpil Im, Andreas Engel, Tilman Schirmer, Benoit Roux, and Daniel J. Müller. "Imaging the Electrostatic Potential of Transmembrane Channels: Atomic Probe Microscopy of OmpF Porin." Biophysical Journal 82.3 (2002): 1667-676. http://dx.doi.org/10.1016/S0006-3495(02)75517-3.

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    KU Libraries
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    Lawrence, KS 66045
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    Contact KU ScholarWorks
    785-864-8983
    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    785-864-8983

    KU Libraries
    1425 Jayhawk Blvd
    Lawrence, KS 66045
    Image Credits
     

     

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