Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain
The anthrax lethal toxin consists of protective antigen (PA) and lethal factor (LF). Understanding both the PA pore formation and LF translocation through the PA pore is crucial to mitigating and perhaps preventing anthrax disease. To better understand the interactions of the LF-PA engagement comple...
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2017-09-01
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doaj-fede19d32f3a48ffa74c7d3b15e737372020-11-25T00:10:11ZengMDPI AGToxins2072-66512017-09-0191029810.3390/toxins9100298toxins9100298Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal DomainAlexandra J. Machen0Narahari Akkaladevi1Caleb Trecazzi2Pierce T. O’Neil3Srayanta Mukherjee4Yifei Qi5Rebecca Dillard6Wonpil Im7Edward P. Gogol8Tommi A. White9Mark T. Fisher10Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USADepartment of Biochemistry, University of Missouri, Columbia, MO 65211, USADepartment of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USADepartment of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USADepartment of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USADepartments of Biological Sciences and Bioengineering, Lehigh University, Bethlehem, PA 18015, USANational Center for Macromolecular Imaging, Baylor University, Houston TX 77030, USADepartments of Biological Sciences and Bioengineering, Lehigh University, Bethlehem, PA 18015, USADepartment of Biological Sciences, University of Missouri-Kansas City, Kansas City, MO 64110, USADepartment of Biochemistry, University of Missouri, Columbia, MO 65211, USADepartment of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USAThe anthrax lethal toxin consists of protective antigen (PA) and lethal factor (LF). Understanding both the PA pore formation and LF translocation through the PA pore is crucial to mitigating and perhaps preventing anthrax disease. To better understand the interactions of the LF-PA engagement complex, the structure of the LFN-bound PA pore solubilized by a lipid nanodisc was examined using cryo-EM. CryoSPARC was used to rapidly sort particle populations of a heterogeneous sample preparation without imposing symmetry, resulting in a refined 17 Å PA pore structure with 3 LFN bound. At pH 7.5, the contributions from the three unstructured LFN lysine-rich tail regions do not occlude the Phe clamp opening. The open Phe clamp suggests that, in this translocation-compromised pH environment, the lysine-rich tails remain flexible and do not interact with the pore lumen region.https://www.mdpi.com/2072-6651/9/10/298anthrax toxinlethal factorprotective antigenpore formationtranslocationnanodisccryo-EMcryoSPARC |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Alexandra J. Machen Narahari Akkaladevi Caleb Trecazzi Pierce T. O’Neil Srayanta Mukherjee Yifei Qi Rebecca Dillard Wonpil Im Edward P. Gogol Tommi A. White Mark T. Fisher |
spellingShingle |
Alexandra J. Machen Narahari Akkaladevi Caleb Trecazzi Pierce T. O’Neil Srayanta Mukherjee Yifei Qi Rebecca Dillard Wonpil Im Edward P. Gogol Tommi A. White Mark T. Fisher Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain Toxins anthrax toxin lethal factor protective antigen pore formation translocation nanodisc cryo-EM cryoSPARC |
author_facet |
Alexandra J. Machen Narahari Akkaladevi Caleb Trecazzi Pierce T. O’Neil Srayanta Mukherjee Yifei Qi Rebecca Dillard Wonpil Im Edward P. Gogol Tommi A. White Mark T. Fisher |
author_sort |
Alexandra J. Machen |
title |
Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain |
title_short |
Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain |
title_full |
Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain |
title_fullStr |
Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain |
title_full_unstemmed |
Asymmetric Cryo-EM Structure of Anthrax Toxin Protective Antigen Pore with Lethal Factor N-Terminal Domain |
title_sort |
asymmetric cryo-em structure of anthrax toxin protective antigen pore with lethal factor n-terminal domain |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2017-09-01 |
description |
The anthrax lethal toxin consists of protective antigen (PA) and lethal factor (LF). Understanding both the PA pore formation and LF translocation through the PA pore is crucial to mitigating and perhaps preventing anthrax disease. To better understand the interactions of the LF-PA engagement complex, the structure of the LFN-bound PA pore solubilized by a lipid nanodisc was examined using cryo-EM. CryoSPARC was used to rapidly sort particle populations of a heterogeneous sample preparation without imposing symmetry, resulting in a refined 17 Å PA pore structure with 3 LFN bound. At pH 7.5, the contributions from the three unstructured LFN lysine-rich tail regions do not occlude the Phe clamp opening. The open Phe clamp suggests that, in this translocation-compromised pH environment, the lysine-rich tails remain flexible and do not interact with the pore lumen region. |
topic |
anthrax toxin lethal factor protective antigen pore formation translocation nanodisc cryo-EM cryoSPARC |
url |
https://www.mdpi.com/2072-6651/9/10/298 |
work_keys_str_mv |
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