Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane

Enteropathogenic Escherichia coli O127 is encapsulated by a protective layer of polysaccharide made of the same strain specific O-antigen as the serotype lipopolysaccharide. Seven genes encoding capsule export functions comprise the group 4 capsule (gfc) operon. Genes gfcE, etk and etp encode homolo...

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Autores principales:	Matthew R. Larson, Kassia Biddle, Adam Gorman, Sarah Boutom, Ilan Rosenshine, Mark A. Saper
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1049d3c053934f9f9eaa72b2163ee366
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spelling	oai:doaj.org-article:1049d3c053934f9f9eaa72b2163ee3662021-11-25T06:10:53ZEscherichia coli O127 group 4 capsule proteins assemble at the outer membrane1932-6203https://doaj.org/article/1049d3c053934f9f9eaa72b2163ee3662021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8592465/?tool=EBIhttps://doaj.org/toc/1932-6203Enteropathogenic Escherichia coli O127 is encapsulated by a protective layer of polysaccharide made of the same strain specific O-antigen as the serotype lipopolysaccharide. Seven genes encoding capsule export functions comprise the group 4 capsule (gfc) operon. Genes gfcE, etk and etp encode homologs of the group 1 capsule secretion system but the upstream gfcABCD genes encode unknown functions specific to group 4 capsule export. We have developed an expression system for the large-scale production of the outer membrane protein GfcD. Contrary to annotations, we find that GfcD is a non-acylated integral membrane protein. Circular dichroism spectroscopy, light-scattering data, and the HHomp server suggested that GfcD is a monomeric β-barrel with 26 β-strands and an internal globular domain. We identified a set of novel protein-protein interactions between GfcB, GfcC, and GfcD, both in vivo and in vitro, and quantified the binding properties with isothermal calorimetry and biolayer interferometry. GfcC and GfcB form a high-affinity heterodimer with a KD near 100 nM. This heterodimer binds to GfcD (KD = 28 μM) significantly better than either GfcB or GfcC alone. These gfc proteins may form a complex at the outer membrane for group 4 capsule secretion or for a yet unknown function.Matthew R. LarsonKassia BiddleAdam GormanSarah BoutomIlan RosenshineMark A. SaperPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Matthew R. Larson Kassia Biddle Adam Gorman Sarah Boutom Ilan Rosenshine Mark A. Saper Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane
description	Enteropathogenic Escherichia coli O127 is encapsulated by a protective layer of polysaccharide made of the same strain specific O-antigen as the serotype lipopolysaccharide. Seven genes encoding capsule export functions comprise the group 4 capsule (gfc) operon. Genes gfcE, etk and etp encode homologs of the group 1 capsule secretion system but the upstream gfcABCD genes encode unknown functions specific to group 4 capsule export. We have developed an expression system for the large-scale production of the outer membrane protein GfcD. Contrary to annotations, we find that GfcD is a non-acylated integral membrane protein. Circular dichroism spectroscopy, light-scattering data, and the HHomp server suggested that GfcD is a monomeric β-barrel with 26 β-strands and an internal globular domain. We identified a set of novel protein-protein interactions between GfcB, GfcC, and GfcD, both in vivo and in vitro, and quantified the binding properties with isothermal calorimetry and biolayer interferometry. GfcC and GfcB form a high-affinity heterodimer with a KD near 100 nM. This heterodimer binds to GfcD (KD = 28 μM) significantly better than either GfcB or GfcC alone. These gfc proteins may form a complex at the outer membrane for group 4 capsule secretion or for a yet unknown function.
format	article
author	Matthew R. Larson Kassia Biddle Adam Gorman Sarah Boutom Ilan Rosenshine Mark A. Saper
author_facet	Matthew R. Larson Kassia Biddle Adam Gorman Sarah Boutom Ilan Rosenshine Mark A. Saper
author_sort	Matthew R. Larson
title	Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane
title_short	Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane
title_full	Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane
title_fullStr	Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane
title_full_unstemmed	Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane
title_sort	escherichia coli o127 group 4 capsule proteins assemble at the outer membrane
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/1049d3c053934f9f9eaa72b2163ee366
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Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane

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