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Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector
BteA, a 69-kDa cytotoxic protein, is a type III secretion system (T3SS) effector in the classical Bordetella, the etiological agents of pertussis and related mammalian respiratory diseases. Currently there is limited information regarding the structure of BteA or its subdomains, and no insight as to...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3559503/ https://www.ncbi.nlm.nih.gov/pubmed/23383256 http://dx.doi.org/10.1371/journal.pone.0055650 |
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author | Guttman, Chen Davidov, Geula Shaked, Hadassa Kolusheva, Sofiya Bitton, Ronit Ganguly, Atish Miller, Jeff F. Chill, Jordan H. Zarivach, Raz |
author_facet | Guttman, Chen Davidov, Geula Shaked, Hadassa Kolusheva, Sofiya Bitton, Ronit Ganguly, Atish Miller, Jeff F. Chill, Jordan H. Zarivach, Raz |
author_sort | Guttman, Chen |
collection | PubMed |
description | BteA, a 69-kDa cytotoxic protein, is a type III secretion system (T3SS) effector in the classical Bordetella, the etiological agents of pertussis and related mammalian respiratory diseases. Currently there is limited information regarding the structure of BteA or its subdomains, and no insight as to the identity of its eukaryotic partners(s) and their modes of interaction with BteA. The mechanisms that lead to BteA dependent cell death also remain elusive. The N-terminal domain of BteA is multifunctional, acting as a docking platform for its cognate chaperone (BtcA) in the bacterium, and targeting the protein to lipid raft microdomains within the eukaryotic host cell. In this study we describe the biochemical and biophysical characteristics of this domain (BteA287) and determine its architecture. We characterize BteA287 as being a soluble and highly stable domain which is rich in alpha helical content. Nuclear magnetic resonance (NMR) experiments combined with size exclusion and analytical ultracentrifugation measurements confirm these observations and reveal BteA287 to be monomeric in nature with a tendency to oligomerize at concentrations above 200 µM. Furthermore, diffusion-NMR demonstrated that the first 31 residues of BteA287 are responsible for the apparent aggregation behavior of BteA287. Light scattering analyses and small angle X-ray scattering experiments reveal a prolate ellipsoidal bi-pyramidal dumb-bell shape. Thus, our biophysical characterization is a first step towards structure determination of the BteA N-terminal domain. |
format | Online Article Text |
id | pubmed-3559503 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-35595032013-02-04 Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector Guttman, Chen Davidov, Geula Shaked, Hadassa Kolusheva, Sofiya Bitton, Ronit Ganguly, Atish Miller, Jeff F. Chill, Jordan H. Zarivach, Raz PLoS One Research Article BteA, a 69-kDa cytotoxic protein, is a type III secretion system (T3SS) effector in the classical Bordetella, the etiological agents of pertussis and related mammalian respiratory diseases. Currently there is limited information regarding the structure of BteA or its subdomains, and no insight as to the identity of its eukaryotic partners(s) and their modes of interaction with BteA. The mechanisms that lead to BteA dependent cell death also remain elusive. The N-terminal domain of BteA is multifunctional, acting as a docking platform for its cognate chaperone (BtcA) in the bacterium, and targeting the protein to lipid raft microdomains within the eukaryotic host cell. In this study we describe the biochemical and biophysical characteristics of this domain (BteA287) and determine its architecture. We characterize BteA287 as being a soluble and highly stable domain which is rich in alpha helical content. Nuclear magnetic resonance (NMR) experiments combined with size exclusion and analytical ultracentrifugation measurements confirm these observations and reveal BteA287 to be monomeric in nature with a tendency to oligomerize at concentrations above 200 µM. Furthermore, diffusion-NMR demonstrated that the first 31 residues of BteA287 are responsible for the apparent aggregation behavior of BteA287. Light scattering analyses and small angle X-ray scattering experiments reveal a prolate ellipsoidal bi-pyramidal dumb-bell shape. Thus, our biophysical characterization is a first step towards structure determination of the BteA N-terminal domain. Public Library of Science 2013-01-30 /pmc/articles/PMC3559503/ /pubmed/23383256 http://dx.doi.org/10.1371/journal.pone.0055650 Text en © 2013 Guttman 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 Guttman, Chen Davidov, Geula Shaked, Hadassa Kolusheva, Sofiya Bitton, Ronit Ganguly, Atish Miller, Jeff F. Chill, Jordan H. Zarivach, Raz Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector |
title | Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector |
title_full | Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector |
title_fullStr | Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector |
title_full_unstemmed | Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector |
title_short | Characterization of the N-Terminal Domain of BteA: A Bordetella Type III Secreted Cytotoxic Effector |
title_sort | characterization of the n-terminal domain of btea: a bordetella type iii secreted cytotoxic effector |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3559503/ https://www.ncbi.nlm.nih.gov/pubmed/23383256 http://dx.doi.org/10.1371/journal.pone.0055650 |
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