Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles

ABSTRACT Outer membrane vesicles (OMV) are spherical membranous structures released from the outer membrane (OM) of Gram-negative bacteria. OMV have been proposed to play several different roles during both pathogenesis and symbiosis. Despite the fact that OMV were described several decades ago, the...

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Autores principales:	Wael Elhenawy, Mykhaylo O. Debelyy, Mario F. Feldman
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/180f8d5603874bfbaf4b2b4ebea6f9fe
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spelling	oai:doaj.org-article:180f8d5603874bfbaf4b2b4ebea6f9fe2021-11-15T15:45:13ZPreferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles10.1128/mBio.00909-142150-7511https://doaj.org/article/180f8d5603874bfbaf4b2b4ebea6f9fe2014-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00909-14https://doaj.org/toc/2150-7511ABSTRACT Outer membrane vesicles (OMV) are spherical membranous structures released from the outer membrane (OM) of Gram-negative bacteria. OMV have been proposed to play several different roles during both pathogenesis and symbiosis. Despite the fact that OMV were described several decades ago, their biogenesis is a poorly characterized process. Whether OMV are produced by an active mechanism or by passive disintegration of the OM is a still matter of controversy. Bacteroides fragilis and Bacteroides thetaiotaomicron are important members of the human microbiota. In this work, we determined and compared the protein compositions of OM and OMV from B. fragilis and B. thetaiotaomicron. SDS-PAGE analysis of both fractions revealed dramatically different protein profiles. Proteomic analysis of OM and OMV in B. fragilis identified more than 40 proteins found exclusively in OMV and more than 30 proteins detectable only in the OM. The OMV-specific proteome showed a high prevalence of glycosidases and proteases, some of which were shown to be active in vitro. Similar results were obtained for B. thetaiotaomicron. Most of the OMV-exclusive proteins were acidic. Based on these results, we propose that these species possess machinery devoted to selectively pack acidic proteins into the OMV. These OMV equipped with hydrolytic enzymes could help in securing nutrients for the benefit of the whole bacterial community present in the microbiota, uncovering a novel function for bacterial OMV. IMPORTANCE The members of genus Bacteroides are key players in the symbiosis between the human host and the gut microbiota. It is known for its ability to degrade a wide variety of glycans that are not substrates for human glycosidases. The cleaved glycans can be utilized by Bacteroides and other microbiota members, resulting in the production of short-chain fatty acids that are beneficial for the host. Although members of the genus Bacteroides are known to secrete different hydrolases, their secretion pathways remain uncharacterized. In this article, we show that B. fragilis and B. thetaiotaomicron preferentially pack a large number of hydrolases in outer membrane vesicles (OMV). Most of these hydrolases are acidic and were detected exclusively in OMV. This suggests the presence of a molecular mechanism in Bacteroides responsible for the selection of OMV proteins based on their charge. We propose that OMV contribute to the establishment and balance of the gut microbiota.Wael ElhenawyMykhaylo O. DebelyyMario F. FeldmanAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 2 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Wael Elhenawy Mykhaylo O. Debelyy Mario F. Feldman Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles
description	ABSTRACT Outer membrane vesicles (OMV) are spherical membranous structures released from the outer membrane (OM) of Gram-negative bacteria. OMV have been proposed to play several different roles during both pathogenesis and symbiosis. Despite the fact that OMV were described several decades ago, their biogenesis is a poorly characterized process. Whether OMV are produced by an active mechanism or by passive disintegration of the OM is a still matter of controversy. Bacteroides fragilis and Bacteroides thetaiotaomicron are important members of the human microbiota. In this work, we determined and compared the protein compositions of OM and OMV from B. fragilis and B. thetaiotaomicron. SDS-PAGE analysis of both fractions revealed dramatically different protein profiles. Proteomic analysis of OM and OMV in B. fragilis identified more than 40 proteins found exclusively in OMV and more than 30 proteins detectable only in the OM. The OMV-specific proteome showed a high prevalence of glycosidases and proteases, some of which were shown to be active in vitro. Similar results were obtained for B. thetaiotaomicron. Most of the OMV-exclusive proteins were acidic. Based on these results, we propose that these species possess machinery devoted to selectively pack acidic proteins into the OMV. These OMV equipped with hydrolytic enzymes could help in securing nutrients for the benefit of the whole bacterial community present in the microbiota, uncovering a novel function for bacterial OMV. IMPORTANCE The members of genus Bacteroides are key players in the symbiosis between the human host and the gut microbiota. It is known for its ability to degrade a wide variety of glycans that are not substrates for human glycosidases. The cleaved glycans can be utilized by Bacteroides and other microbiota members, resulting in the production of short-chain fatty acids that are beneficial for the host. Although members of the genus Bacteroides are known to secrete different hydrolases, their secretion pathways remain uncharacterized. In this article, we show that B. fragilis and B. thetaiotaomicron preferentially pack a large number of hydrolases in outer membrane vesicles (OMV). Most of these hydrolases are acidic and were detected exclusively in OMV. This suggests the presence of a molecular mechanism in Bacteroides responsible for the selection of OMV proteins based on their charge. We propose that OMV contribute to the establishment and balance of the gut microbiota.
format	article
author	Wael Elhenawy Mykhaylo O. Debelyy Mario F. Feldman
author_facet	Wael Elhenawy Mykhaylo O. Debelyy Mario F. Feldman
author_sort	Wael Elhenawy
title	Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles
title_short	Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles
title_full	Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles
title_fullStr	Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles
title_full_unstemmed	Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles
title_sort	preferential packing of acidic glycosidases and proteases into <italic toggle="yes">bacteroides</italic> outer membrane vesicles
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/180f8d5603874bfbaf4b2b4ebea6f9fe
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Preferential Packing of Acidic Glycosidases and Proteases into <italic toggle="yes">Bacteroides</italic> Outer Membrane Vesicles

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