Technical Report abstract
Nature Structural & Molecular Biology 15, 754 - 760 (2008)
Published online: 22 June 2008 | doi:10.1038/nsmb.1442
GroEL as a molecular scaffold for structural analysis of the anthrax toxin pore
Hiroo Katayama1,5, Blythe E Janowiak2,5, Marek Brzozowski1, Jordan Juryck1, Scott Falke3, Edward P Gogol4, R John Collier2 & Mark T Fisher1
Abstract
We analyzed the 440-kDa transmembrane pore formed by the protective antigen (PA) moiety of anthrax toxin in the presence of GroEL by negative-stain electron microscopy. GroEL binds both the heptameric PA prepore and the PA pore. The latter interaction retards aggregation of the pore, prolonging its insertion-competent state. Two populations of unaggregated pores were visible: GroEL-bound pores and unbound pores. This allowed two virtually identical structures to be reconstructed, at 25-Å and 28-Å resolution, respectively. The structures were mushroom-shaped objects with a 125-Å-diameter cap and a 100-Å-long stem, consistent with earlier biochemical data. Thus, GroEL provides a platform for obtaining initial glimpses of a membrane protein structure in the absence of lipids or detergents and can function as a scaffold for higher-resolution structural analysis of the PA pore.
- Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, 3901 Rainbow Blvd., Kansas Life Sciences Innovation Center Building, Kansas City, Kansas 66160, USA.
- Department of Microbiology and Molecular Genetics, Harvard Medical School, Alpert Building, Boston, Massachusetts 02115, USA.
- Department of Biology, William Jewell College, 500 College Hill, Liberty, Missouri 64068-1896, USA.
- School of Biological Sciences, University of Missouri-Kansas City, Kansas City, Missouri 64100, USA.
- These authors contributed equally to this work.
Correspondence to: Mark T Fisher1 e-mail: mfisher1@kumc.edu
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