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MICROBIOLOGY
A model of anthrax toxin lethal factor bound to protective antigen

D. Borden Lacy ^*, Henry C. Lin ^*, Roman A. Melnyk ^*, Ora Schueler-Furman , Laura Reither , Kristina Cunningham ^* , David Baker ^, ¶, and R. John Collier ^*, ||

^*Department of Microbiology and Molecular Genetics and Graduate Group of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115; and Department of Biochemistry and ^¶Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195

Contributed by R. John Collier, September 21, 2005

Anthrax toxin is made up of three proteins: the edema factor (EF), lethal factor (LF) enzymes, and the multifunctional protective antigen (PA). Proteolytically activated PA heptamerizes, binds the EF/LF enzymes, and forms a pore that allows for EF/LF passage into host cells. Using directed mutagenesis, we identified three LF-PA contact points defined by a specific disulfide crosslink and two pairs of complementary charge-reversal mutations. These contact points were consistent with the lowest energy LF-PA complex found by using Rosetta protein-protein docking. These results illustrate how biochemical and computational methods can be combined to produce reliable models of large complexes. The model shows that EF and LF bind through a highly electrostatic interface, with their flexible N-terminal region positioned at the entrance of the heptameric PA pore and thus poised to initiate translocation in an N- to C-terminal direction.

computation | docking | electrostatic

Conflict of interest statement: R.J.C. holds equity in PharmAthene, Inc., a company engaged in developing countermeasures to biothreat agents, including Bacillus anthracis.

Abbreviations: EF, edema factor; LF, lethal factor; PA, protective antigen; rmsd, rms deviation.

Present address: Wyeth Research, Cambridge, MA 02140.

^|| To whom correspondence should be addressed. E-mail: john_collier{at}hms.harvard.edu.

© 2005 by The National Academy of Sciences of the USA

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