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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Public Library of Science (PLoS)
2021-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0259900&type=printable |
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| 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 |
| collection | DOAJ |
| 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 |
| id | doaj-art-333b0ce93bb94bbeb53fe28f8142ebd6 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj-art-333b0ce93bb94bbeb53fe28f8142ebd62025-01-21T05:31:29ZengPublic Library of Science (PLoS)PLoS ONE1932-62032021-01-011611e025990010.1371/journal.pone.0259900Escherichia coli O127 group 4 capsule proteins assemble at the outer membrane.Matthew R LarsonKassia BiddleAdam GormanSarah BoutomIlan RosenshineMark A SaperEnteropathogenic 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0259900&type=printable |
| spellingShingle | 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. PLoS ONE |
| title | 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_short | 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 |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0259900&type=printable |
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