Structural Basis for the Unfolding of Anthrax Lethal Factor by Protective Antigen Oligomers

Overview

Journal Nat Struct Mol Biol

Specialties Cell Biology
Molecular Biology

Date 2010 Nov 2

PMID 21037566

Citations 75

Authors

Geoffrey K Feld

Katie L Thoren

Alexander F Kintzer

Harry J Sterling

Iok I Tang

Shoshana G Greenberg

Evan R Williams

Bryan A Krantz

Affiliations

Soon will be listed here.

Abstract

The protein transporter anthrax lethal toxin is composed of protective antigen (PA), a transmembrane translocase, and lethal factor (LF), a cytotoxic enzyme. After its assembly into holotoxin complexes, PA forms an oligomeric channel that unfolds LF and translocates it into the host cell. We report the crystal structure of the core of a lethal toxin complex to 3.1-Å resolution; the structure contains a PA octamer bound to four LF PA-binding domains (LF(N)). The first α-helix and β-strand of each LF(N) unfold and dock into a deep amphipathic cleft on the surface of the PA octamer, which we call the α clamp. The α clamp possesses nonspecific polypeptide binding activity and is functionally relevant to efficient holotoxin assembly, PA octamer formation, and LF unfolding and translocation. This structure provides insight into the mechanism of translocation-coupled protein unfolding.

Citing Articles

Targeting the Inside of Cells with Biologicals: Toxin Routes in a Therapeutic Context.

Ruschig M, Marschall A BioDrugs. 2023; 37(2):181-203.

PMID: 36729328 PMC: 9893211. DOI: 10.1007/s40259-023-00580-y.

Solution Structures of Protective Antigen Proteins Using Small Angle Neutron Scattering and Protective Antigen 63 Ion Channel Formation Kinetics.

Michelman-Ribeiro A, Rubinson K, Silin V, Kasianowicz J Toxins (Basel). 2021; 13(12).

PMID: 34941724 PMC: 8708185. DOI: 10.3390/toxins13120888.

Thermodynamic Stability Is a Strong Predictor for the Delivery of DARPins to the Cytosol via Anthrax Toxin.

Becker L, Singh Badwal J, Brandl F, Verdurmen W, Pluckthun A Pharmaceutics. 2021; 13(8).

PMID: 34452246 PMC: 8401532. DOI: 10.3390/pharmaceutics13081285.

Anthrax toxin translocation complex reveals insight into the lethal factor unfolding and refolding mechanism.

Machen A, Fisher M, Freudenthal B Sci Rep. 2021; 11(1):13038.

PMID: 34158520 PMC: 8219829. DOI: 10.1038/s41598-021-91596-3.

Natural and Designed Toxins for Precise Therapy: Modern Approaches in Experimental Oncology.

Shilova O, Shramova E, Proshkina G, Deyev S Int J Mol Sci. 2021; 22(9).

PMID: 34067057 PMC: 8124712. DOI: 10.3390/ijms22094975.

References

Duesbery N, Webb C, Leppla S, Gordon V, Klimpel K, Copeland T . Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor. Science. 1998; 280(5364):734-7. DOI: 10.1126/science.280.5364.734. View

Melnyk R, Hewitt K, Lacy D, Lin H, Gessner C, Li S . Structural determinants for the binding of anthrax lethal factor to oligomeric protective antigen. J Biol Chem. 2005; 281(3):1630-5. DOI: 10.1074/jbc.M511164200. View

Meador W, Means A, Quiocho F . Modulation of calmodulin plasticity in molecular recognition on the basis of x-ray structures. Science. 1993; 262(5140):1718-21. DOI: 10.1126/science.8259515. View

Cheng Y . Toward an atomic model of the 26S proteasome. Curr Opin Struct Biol. 2009; 19(2):203-8. PMC: 2743420. DOI: 10.1016/j.sbi.2009.02.004. View

Wickner W, Schekman R . Protein translocation across biological membranes. Science. 2005; 310(5753):1452-6. DOI: 10.1126/science.1113752. View

Clackson T, Wells J . A hot spot of binding energy in a hormone-receptor interface. Science. 1995; 267(5196):383-6. DOI: 10.1126/science.7529940. View

Sauer R, Bolon D, Burton B, Burton R, Flynn J, Grant R . Sculpting the proteome with AAA(+) proteases and disassembly machines. Cell. 2004; 119(1):9-18. PMC: 2717008. DOI: 10.1016/j.cell.2004.09.020. View

Krantz B, Melnyk R, Zhang S, Juris S, Lacy D, Wu Z . A phenylalanine clamp catalyzes protein translocation through the anthrax toxin pore. Science. 2005; 309(5735):777-81. PMC: 1815389. DOI: 10.1126/science.1113380. View

Milne J, Furlong D, Hanna P, Wall J, Collier R . Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J Biol Chem. 1994; 269(32):20607-12. View

10.

Katayama H, Janowiak B, Brzozowski M, Juryck J, Falke S, Gogol E . GroEL as a molecular scaffold for structural analysis of the anthrax toxin pore. Nat Struct Mol Biol. 2008; 15(7):754-60. PMC: 2504863. DOI: 10.1038/nsmb.1442. View

11.

Arora N, Leppla S . Residues 1-254 of anthrax toxin lethal factor are sufficient to cause cellular uptake of fused polypeptides. J Biol Chem. 1993; 268(5):3334-41. View

12.

Wang Z, Feng H, Landry S, Maxwell J, Gierasch L . Basis of substrate binding by the chaperonin GroEL. Biochemistry. 1999; 38(39):12537-46. DOI: 10.1021/bi991070p. View

13.

Petosa C, Collier R, Klimpel K, Leppla S, Liddington R . Crystal structure of the anthrax toxin protective antigen. Nature. 1997; 385(6619):833-8. DOI: 10.1038/385833a0. View

14.

Pettersen E, Goddard T, Huang C, Couch G, Greenblatt D, Meng E . UCSF Chimera--a visualization system for exploratory research and analysis. J Comput Chem. 2004; 25(13):1605-12. DOI: 10.1002/jcc.20084. View

15.

Milne J, Blanke S, Hanna P, Collier R . Protective antigen-binding domain of anthrax lethal factor mediates translocation of a heterologous protein fused to its amino- or carboxy-terminus. Mol Microbiol. 1995; 15(4):661-6. DOI: 10.1111/j.1365-2958.1995.tb02375.x. View

16.

Kintzer A, Sterling H, Tang I, Abdul-Gader A, Miles A, Wallace B . Role of the protective antigen octamer in the molecular mechanism of anthrax lethal toxin stabilization in plasma. J Mol Biol. 2010; 399(5):741-58. PMC: 2892534. DOI: 10.1016/j.jmb.2010.04.041. View

17.

Thoren K, Worden E, Yassif J, Krantz B . Lethal factor unfolding is the most force-dependent step of anthrax toxin translocation. Proc Natl Acad Sci U S A. 2009; 106(51):21555-60. PMC: 2779827. DOI: 10.1073/pnas.0905880106. View

18.

Lum R, Niggemann M, Glover J . Peptide and protein binding in the axial channel of Hsp104. Insights into the mechanism of protein unfolding. J Biol Chem. 2008; 283(44):30139-50. PMC: 2662077. DOI: 10.1074/jbc.M804849200. View

19.

Krantz B, Finkelstein A, Collier R . Protein translocation through the anthrax toxin transmembrane pore is driven by a proton gradient. J Mol Biol. 2005; 355(5):968-79. DOI: 10.1016/j.jmb.2005.11.030. View

20.

Davis I, Leaver-Fay A, Chen V, Block J, Kapral G, Wang X . MolProbity: all-atom contacts and structure validation for proteins and nucleic acids. Nucleic Acids Res. 2007; 35(Web Server issue):W375-83. PMC: 1933162. DOI: 10.1093/nar/gkm216. View