Imaging of reconstituted biological channels at molecular resolution by atomic force microscopy.
Using atomic force microscopy (AFM), we obtained high-resolution surface images of the bacterial outer membrane channels Escherichia coli OmpF porin and Bordetella pertussis porin that were reconstituted in artificial bilayer membranes as two-dimensional crystalline arrays. These porins were chosen because they are among the most extensively studied proteins of this type and are known for their well-defined crystalline nature in the native membrane. Such reconstituted membrane proteins are ideal specimens to assess the suitability and resolution of AFM for imaging biomembranes and associated proteins. Although OmpF porin often showed a mixed pattern of rectangular and hexagonal arrays with approximately 8.4 x 9.8- and approximately 7.2-nm-spacings, respectively, B. pertussis porin showed mostly a rectangular pattern with an approximately 7.9 x 13.8-nm spacing. The packing patterns of the E. coli OmpF porin in the membrane are very close to those found in electron-microscopic studies. When B. pertussis porin was imaged in a buffer solution, its trimeric subunits were apparently resolved, and the surface of each monomer revealed beadlike structures. This is the first report of such a high-resolution structural analysis of B. pertussis porin by any imaging method. We also imaged the lipid bilayer itself as an internal control for imaging and to further ascertain the resolution. Individual polar head groups of bilayer lipid molecules were resolved, suggesting the intrinsic resolution of AFM for bioimaging.
Lal
R
R
Department of Medicine, University of Chicago, Illinois 60637.
Kim
H
H
Garavito
R M
RM
Arnsdorf
M F
MF
D001886:Bordetella pertussis / Q000378:metabolism; D004134:Dimyristoylphosphatidylcholine; D004926:Escherichia coli / Q000378:metabolism; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D008853:Microscopy / Q000379:methods*; D016252:Microscopy, Scanning Tunneling; D018272:Porins / Q000378:metabolism / Q000648:ultrastructure*
D008051:Lipid Bilayers; D008567:Membranes, Artificial; D018272:Porins; D004134:Dimyristoylphosphatidylcholine