Structure of EspB, a secreted substrate of the ESX-1 secretion system of Mycobacterium tuberculosis

Natalia Korotkova; Jérémie Piton; Jonathan M. Wagner; Stefanie Boy-Röttger; Aleksandre Japaridze; Timothy J. Evans; Stewart T. Cole; Florence Pojer; Konstantin V. Korotkov

doi:10.1016/j.jsb.2015.06.003

Structure of EspB, a secreted substrate of the ESX-1 secretion system of Mycobacterium tuberculosis

Natalia Korotkova, Jérémie Piton, Jonathan M. Wagner, Stefanie Boy-Röttger, Aleksandre Japaridze, Timothy J. Evans, Stewart T. Cole, Florence Pojer, Konstantin V. Korotkov

Research output: Contribution to journal › Article › peer-review

33 Citations (SciVal)

Abstract

Mycobacterium tuberculosis secretes multiple virulence factors during infection via the general Sec and Tat pathways, and via specialized ESX secretion systems, also referred to as type VII secretion systems. The ESX-1 secretion system is an important virulence determinant because deletion of ESX-1 leads to attenuation of M. tuberculosis. ESX-1 secreted protein B (EspB) contains putative PE (Pro-Glu) and PPE (Pro-Pro-Glu) domains, and a C-terminal domain, which is processed by MycP₁ protease during secretion. We determined the crystal structure of PE-PPE domains of EspB, which represents an all-helical, elongated molecule closely resembling the structure of the PE25-PPE41 heterodimer despite limited sequence similarity. Also, we determined the structure of full-length EspB, which does not have interpretable electron density for the C-terminal domain confirming that it is largely disordered. Comparative analysis of EspB in cell lysate and culture filtrates of M. tuberculosis revealed that mature secreted EspB forms oligomers. Electron microscopy analysis showed that the N-terminal fragment of EspB forms donut-shaped particles. These data provide a rationale for the future investigation of EspB's role in M. tuberculosis pathogenesis.

Original language	English
Pages (from-to)	236-244
Number of pages	9
Journal	Journal of Structural Biology
Volume	191
Issue number	2
DOIs	https://doi.org/10.1016/j.jsb.2015.06.003
State	Published - Aug 1 2015

Bibliographical note

Funding Information:
Authors thank staff members of Southeast Regional Collaborative Access Team (SER-CAT) at the Advanced Photon Source, Argonne National Laboratory, for assistance during data collection. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences , under Contract No. W-31-109-Eng-38 . We thank the EPFL Bioelectron Microscopy and Protein Crystallography Core Facilities, and the Swiss Light Source for beam time and excellent support. We acknowledge the University of Kentucky Protein Analytical Core that is supported by NIH/NIGMS grant P30GM110787 . This study was supported in part by NIH/NIGMS grant P20GM103486 to K.V.K. This study received funding from the Swiss National Science Foundation to S.T.C. ( 31003A-140778 ).

Publisher Copyright:
© 2015 Elsevier Inc.

Keywords

ESX
PE domain
PPE domain
Type VII secretion system

ASJC Scopus subject areas

Structural Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.jsb.2015.06.003

Cite this

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title = "Structure of EspB, a secreted substrate of the ESX-1 secretion system of Mycobacterium tuberculosis",

abstract = "Mycobacterium tuberculosis secretes multiple virulence factors during infection via the general Sec and Tat pathways, and via specialized ESX secretion systems, also referred to as type VII secretion systems. The ESX-1 secretion system is an important virulence determinant because deletion of ESX-1 leads to attenuation of M. tuberculosis. ESX-1 secreted protein B (EspB) contains putative PE (Pro-Glu) and PPE (Pro-Pro-Glu) domains, and a C-terminal domain, which is processed by MycP1 protease during secretion. We determined the crystal structure of PE-PPE domains of EspB, which represents an all-helical, elongated molecule closely resembling the structure of the PE25-PPE41 heterodimer despite limited sequence similarity. Also, we determined the structure of full-length EspB, which does not have interpretable electron density for the C-terminal domain confirming that it is largely disordered. Comparative analysis of EspB in cell lysate and culture filtrates of M. tuberculosis revealed that mature secreted EspB forms oligomers. Electron microscopy analysis showed that the N-terminal fragment of EspB forms donut-shaped particles. These data provide a rationale for the future investigation of EspB's role in M. tuberculosis pathogenesis.",

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author = "Natalia Korotkova and J{\'e}r{\'e}mie Piton and Wagner, {Jonathan M.} and Stefanie Boy-R{\"o}ttger and Aleksandre Japaridze and Evans, {Timothy J.} and Cole, {Stewart T.} and Florence Pojer and Korotkov, {Konstantin V.}",

note = "Funding Information: Authors thank staff members of Southeast Regional Collaborative Access Team (SER-CAT) at the Advanced Photon Source, Argonne National Laboratory, for assistance during data collection. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences , under Contract No. W-31-109-Eng-38 . We thank the EPFL Bioelectron Microscopy and Protein Crystallography Core Facilities, and the Swiss Light Source for beam time and excellent support. We acknowledge the University of Kentucky Protein Analytical Core that is supported by NIH/NIGMS grant P30GM110787 . This study was supported in part by NIH/NIGMS grant P20GM103486 to K.V.K. This study received funding from the Swiss National Science Foundation to S.T.C. ( 31003A-140778 ). Publisher Copyright: {\textcopyright} 2015 Elsevier Inc.",

year = "2015",

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doi = "10.1016/j.jsb.2015.06.003",

language = "English",

volume = "191",

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AU - Korotkova, Natalia

AU - Piton, Jérémie

AU - Wagner, Jonathan M.

AU - Boy-Röttger, Stefanie

AU - Japaridze, Aleksandre

AU - Evans, Timothy J.

AU - Cole, Stewart T.

AU - Pojer, Florence

AU - Korotkov, Konstantin V.

N1 - Funding Information: Authors thank staff members of Southeast Regional Collaborative Access Team (SER-CAT) at the Advanced Photon Source, Argonne National Laboratory, for assistance during data collection. Use of the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences , under Contract No. W-31-109-Eng-38 . We thank the EPFL Bioelectron Microscopy and Protein Crystallography Core Facilities, and the Swiss Light Source for beam time and excellent support. We acknowledge the University of Kentucky Protein Analytical Core that is supported by NIH/NIGMS grant P30GM110787 . This study was supported in part by NIH/NIGMS grant P20GM103486 to K.V.K. This study received funding from the Swiss National Science Foundation to S.T.C. ( 31003A-140778 ). Publisher Copyright: © 2015 Elsevier Inc.

PY - 2015/8/1

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N2 - Mycobacterium tuberculosis secretes multiple virulence factors during infection via the general Sec and Tat pathways, and via specialized ESX secretion systems, also referred to as type VII secretion systems. The ESX-1 secretion system is an important virulence determinant because deletion of ESX-1 leads to attenuation of M. tuberculosis. ESX-1 secreted protein B (EspB) contains putative PE (Pro-Glu) and PPE (Pro-Pro-Glu) domains, and a C-terminal domain, which is processed by MycP1 protease during secretion. We determined the crystal structure of PE-PPE domains of EspB, which represents an all-helical, elongated molecule closely resembling the structure of the PE25-PPE41 heterodimer despite limited sequence similarity. Also, we determined the structure of full-length EspB, which does not have interpretable electron density for the C-terminal domain confirming that it is largely disordered. Comparative analysis of EspB in cell lysate and culture filtrates of M. tuberculosis revealed that mature secreted EspB forms oligomers. Electron microscopy analysis showed that the N-terminal fragment of EspB forms donut-shaped particles. These data provide a rationale for the future investigation of EspB's role in M. tuberculosis pathogenesis.

AB - Mycobacterium tuberculosis secretes multiple virulence factors during infection via the general Sec and Tat pathways, and via specialized ESX secretion systems, also referred to as type VII secretion systems. The ESX-1 secretion system is an important virulence determinant because deletion of ESX-1 leads to attenuation of M. tuberculosis. ESX-1 secreted protein B (EspB) contains putative PE (Pro-Glu) and PPE (Pro-Pro-Glu) domains, and a C-terminal domain, which is processed by MycP1 protease during secretion. We determined the crystal structure of PE-PPE domains of EspB, which represents an all-helical, elongated molecule closely resembling the structure of the PE25-PPE41 heterodimer despite limited sequence similarity. Also, we determined the structure of full-length EspB, which does not have interpretable electron density for the C-terminal domain confirming that it is largely disordered. Comparative analysis of EspB in cell lysate and culture filtrates of M. tuberculosis revealed that mature secreted EspB forms oligomers. Electron microscopy analysis showed that the N-terminal fragment of EspB forms donut-shaped particles. These data provide a rationale for the future investigation of EspB's role in M. tuberculosis pathogenesis.

KW - ESX

KW - PE domain

KW - PPE domain

KW - Type VII secretion system

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Structure of EspB, a secreted substrate of the ESX-1 secretion system of Mycobacterium tuberculosis

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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