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The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1

Macrophage-expressed gene 1 (MPEG1/Perforin-2) is a perforin-like protein that functions within the phagolysosome to damage engulfed microbes. MPEG1 is thought to form pores in target membranes, however, its mode of action remains unknown. We use cryo-Electron Microscopy (cryo-EM) to determine the 2...

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Autores principales: Pang, Siew Siew, Bayly-Jones, Charles, Radjainia, Mazdak, Spicer, Bradley A., Law, Ruby H. P., Hodel, Adrian W., Parsons, Edward S., Ekkel, Susan M., Conroy, Paul J., Ramm, Georg, Venugopal, Hariprasad, Bird, Phillip I., Hoogenboom, Bart W., Voskoboinik, Ilia, Gambin, Yann, Sierecki, Emma, Dunstone, Michelle A., Whisstock, James C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6753088/
https://www.ncbi.nlm.nih.gov/pubmed/31537793
http://dx.doi.org/10.1038/s41467-019-12279-2
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author Pang, Siew Siew
Bayly-Jones, Charles
Radjainia, Mazdak
Spicer, Bradley A.
Law, Ruby H. P.
Hodel, Adrian W.
Parsons, Edward S.
Ekkel, Susan M.
Conroy, Paul J.
Ramm, Georg
Venugopal, Hariprasad
Bird, Phillip I.
Hoogenboom, Bart W.
Voskoboinik, Ilia
Gambin, Yann
Sierecki, Emma
Dunstone, Michelle A.
Whisstock, James C.
author_facet Pang, Siew Siew
Bayly-Jones, Charles
Radjainia, Mazdak
Spicer, Bradley A.
Law, Ruby H. P.
Hodel, Adrian W.
Parsons, Edward S.
Ekkel, Susan M.
Conroy, Paul J.
Ramm, Georg
Venugopal, Hariprasad
Bird, Phillip I.
Hoogenboom, Bart W.
Voskoboinik, Ilia
Gambin, Yann
Sierecki, Emma
Dunstone, Michelle A.
Whisstock, James C.
author_sort Pang, Siew Siew
collection PubMed
description Macrophage-expressed gene 1 (MPEG1/Perforin-2) is a perforin-like protein that functions within the phagolysosome to damage engulfed microbes. MPEG1 is thought to form pores in target membranes, however, its mode of action remains unknown. We use cryo-Electron Microscopy (cryo-EM) to determine the 2.4 Å structure of a hexadecameric assembly of MPEG1 that displays the expected features of a soluble prepore complex. We further discover that MPEG1 prepore-like assemblies can be induced to perforate membranes through acidification, such as would occur within maturing phagolysosomes. We next solve the 3.6 Å cryo-EM structure of MPEG1 in complex with liposomes. These data reveal that a multi-vesicular body of 12 kDa (MVB12)-associated β-prism (MABP) domain binds membranes such that the pore-forming machinery of MPEG1 is oriented away from the bound membrane. This unexpected mechanism of membrane interaction suggests that MPEG1 remains bound to the phagolysosome membrane while simultaneously forming pores in engulfed bacterial targets.
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spelling pubmed-67530882019-09-23 The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1 Pang, Siew Siew Bayly-Jones, Charles Radjainia, Mazdak Spicer, Bradley A. Law, Ruby H. P. Hodel, Adrian W. Parsons, Edward S. Ekkel, Susan M. Conroy, Paul J. Ramm, Georg Venugopal, Hariprasad Bird, Phillip I. Hoogenboom, Bart W. Voskoboinik, Ilia Gambin, Yann Sierecki, Emma Dunstone, Michelle A. Whisstock, James C. Nat Commun Article Macrophage-expressed gene 1 (MPEG1/Perforin-2) is a perforin-like protein that functions within the phagolysosome to damage engulfed microbes. MPEG1 is thought to form pores in target membranes, however, its mode of action remains unknown. We use cryo-Electron Microscopy (cryo-EM) to determine the 2.4 Å structure of a hexadecameric assembly of MPEG1 that displays the expected features of a soluble prepore complex. We further discover that MPEG1 prepore-like assemblies can be induced to perforate membranes through acidification, such as would occur within maturing phagolysosomes. We next solve the 3.6 Å cryo-EM structure of MPEG1 in complex with liposomes. These data reveal that a multi-vesicular body of 12 kDa (MVB12)-associated β-prism (MABP) domain binds membranes such that the pore-forming machinery of MPEG1 is oriented away from the bound membrane. This unexpected mechanism of membrane interaction suggests that MPEG1 remains bound to the phagolysosome membrane while simultaneously forming pores in engulfed bacterial targets. Nature Publishing Group UK 2019-09-19 /pmc/articles/PMC6753088/ /pubmed/31537793 http://dx.doi.org/10.1038/s41467-019-12279-2 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Pang, Siew Siew
Bayly-Jones, Charles
Radjainia, Mazdak
Spicer, Bradley A.
Law, Ruby H. P.
Hodel, Adrian W.
Parsons, Edward S.
Ekkel, Susan M.
Conroy, Paul J.
Ramm, Georg
Venugopal, Hariprasad
Bird, Phillip I.
Hoogenboom, Bart W.
Voskoboinik, Ilia
Gambin, Yann
Sierecki, Emma
Dunstone, Michelle A.
Whisstock, James C.
The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1
title The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1
title_full The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1
title_fullStr The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1
title_full_unstemmed The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1
title_short The cryo-EM structure of the acid activatable pore-forming immune effector Macrophage-expressed gene 1
title_sort cryo-em structure of the acid activatable pore-forming immune effector macrophage-expressed gene 1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6753088/
https://www.ncbi.nlm.nih.gov/pubmed/31537793
http://dx.doi.org/10.1038/s41467-019-12279-2
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