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Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins

The cell envelope of mycobacteria, a group of Gram positive bacteria, is composed of a plasma membrane and a Gram-negative-like outer membrane containing mycolic acids. In addition, the surface of the mycobacteria is coated with an ill-characterized layer of extractable, non-covalently linked glycan...

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Autores principales: Sani, Musa, Houben, Edith N. G., Geurtsen, Jeroen, Pierson, Jason, de Punder, Karin, van Zon, Maaike, Wever, Brigitte, Piersma, Sander R., Jiménez, Connie R., Daffé, Mamadou, Appelmelk, Ben J., Bitter, Wilbert, van der Wel, Nicole, Peters, Peter J.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2832766/
https://www.ncbi.nlm.nih.gov/pubmed/20221442
http://dx.doi.org/10.1371/journal.ppat.1000794
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author Sani, Musa
Houben, Edith N. G.
Geurtsen, Jeroen
Pierson, Jason
de Punder, Karin
van Zon, Maaike
Wever, Brigitte
Piersma, Sander R.
Jiménez, Connie R.
Daffé, Mamadou
Appelmelk, Ben J.
Bitter, Wilbert
van der Wel, Nicole
Peters, Peter J.
author_facet Sani, Musa
Houben, Edith N. G.
Geurtsen, Jeroen
Pierson, Jason
de Punder, Karin
van Zon, Maaike
Wever, Brigitte
Piersma, Sander R.
Jiménez, Connie R.
Daffé, Mamadou
Appelmelk, Ben J.
Bitter, Wilbert
van der Wel, Nicole
Peters, Peter J.
author_sort Sani, Musa
collection PubMed
description The cell envelope of mycobacteria, a group of Gram positive bacteria, is composed of a plasma membrane and a Gram-negative-like outer membrane containing mycolic acids. In addition, the surface of the mycobacteria is coated with an ill-characterized layer of extractable, non-covalently linked glycans, lipids and proteins, collectively known as the capsule, whose occurrence is a matter of debate. By using plunge freezing cryo-electron microscopy technique, we were able to show that pathogenic mycobacteria produce a thick capsule, only present when the cells were grown under unperturbed conditions and easily removed by mild detergents. This detergent-labile capsule layer contains arabinomannan, α-glucan and oligomannosyl-capped glycolipids. Further immunogenic and proteomic analyses revealed that Mycobacterium marinum capsule contains high amounts of proteins that are secreted via the ESX-1 pathway. Finally, cell infection experiments demonstrated the importance of the capsule for binding to cells and dampening of pro-inflammatory cytokine response. Together, these results show a direct visualization of the mycobacterial capsular layer as a labile structure that contains ESX-1-secreted proteins.
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spelling pubmed-28327662010-03-10 Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins Sani, Musa Houben, Edith N. G. Geurtsen, Jeroen Pierson, Jason de Punder, Karin van Zon, Maaike Wever, Brigitte Piersma, Sander R. Jiménez, Connie R. Daffé, Mamadou Appelmelk, Ben J. Bitter, Wilbert van der Wel, Nicole Peters, Peter J. PLoS Pathog Research Article The cell envelope of mycobacteria, a group of Gram positive bacteria, is composed of a plasma membrane and a Gram-negative-like outer membrane containing mycolic acids. In addition, the surface of the mycobacteria is coated with an ill-characterized layer of extractable, non-covalently linked glycans, lipids and proteins, collectively known as the capsule, whose occurrence is a matter of debate. By using plunge freezing cryo-electron microscopy technique, we were able to show that pathogenic mycobacteria produce a thick capsule, only present when the cells were grown under unperturbed conditions and easily removed by mild detergents. This detergent-labile capsule layer contains arabinomannan, α-glucan and oligomannosyl-capped glycolipids. Further immunogenic and proteomic analyses revealed that Mycobacterium marinum capsule contains high amounts of proteins that are secreted via the ESX-1 pathway. Finally, cell infection experiments demonstrated the importance of the capsule for binding to cells and dampening of pro-inflammatory cytokine response. Together, these results show a direct visualization of the mycobacterial capsular layer as a labile structure that contains ESX-1-secreted proteins. Public Library of Science 2010-03-05 /pmc/articles/PMC2832766/ /pubmed/20221442 http://dx.doi.org/10.1371/journal.ppat.1000794 Text en Sani et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Sani, Musa
Houben, Edith N. G.
Geurtsen, Jeroen
Pierson, Jason
de Punder, Karin
van Zon, Maaike
Wever, Brigitte
Piersma, Sander R.
Jiménez, Connie R.
Daffé, Mamadou
Appelmelk, Ben J.
Bitter, Wilbert
van der Wel, Nicole
Peters, Peter J.
Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins
title Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins
title_full Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins
title_fullStr Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins
title_full_unstemmed Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins
title_short Direct Visualization by Cryo-EM of the Mycobacterial Capsular Layer: A Labile Structure Containing ESX-1-Secreted Proteins
title_sort direct visualization by cryo-em of the mycobacterial capsular layer: a labile structure containing esx-1-secreted proteins
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2832766/
https://www.ncbi.nlm.nih.gov/pubmed/20221442
http://dx.doi.org/10.1371/journal.ppat.1000794
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