Proteomic characterization of a natural host-pathogen interaction: repertoire of in vivo expressed bacterial and host surface-associated proteins

Megan A Rees, Oded Kleifeld, Paul K Crellin, Bosco Kwan-Chung Ho, Timothy P Stinear, Alexander Ian Smith, Ross L Coppel

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Interactions between a host and a bacterial pathogen are mediated by cross-talk between molecules present on, or secreted by, pathogens and host binding-molecules. Identifying proteins involved at this interface would provide substantial insights into this interaction. While numerous studies have examined in vitro models of infection at the level of transcriptional change and proteomic profiling, there is virtually no information available on naturally occurring host-pathogen interactions in vivo. We employed membrane shaving to identify peptide fragments cleaved from surface-expressed bacterial proteins and also detected proteins originating from the infected host. We optimised this technique for media-cultured Corynebacterium pseudotuberculosis, a sheep pathogen, revealing a set of 247 surface proteins. We then studied a natural host-pathogen interaction, by performing membrane shaving on C. pseudotuberculosis harvested directly from naturally infected sheep lymph nodes. Thirty-one bacterial surface proteins were identified including thirteen not identified in culture media, suggesting a different surface protein repertoire is expressed in this hostile environment. Forty-nine host proteins were identified including immune mediators and antimicrobial peptides such as cathelicidin. This novel application of proteolytic shaving has documented sets of host and pathogen proteins present at the bacterial surface in an infection of the native host.
Original languageEnglish
Pages (from-to)120 - 132
Number of pages13
JournalJournal of Proteome Research
Volume14
Issue number1
DOIs
Publication statusPublished - 2015

Cite this

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title = "Proteomic characterization of a natural host-pathogen interaction: repertoire of in vivo expressed bacterial and host surface-associated proteins",
abstract = "Interactions between a host and a bacterial pathogen are mediated by cross-talk between molecules present on, or secreted by, pathogens and host binding-molecules. Identifying proteins involved at this interface would provide substantial insights into this interaction. While numerous studies have examined in vitro models of infection at the level of transcriptional change and proteomic profiling, there is virtually no information available on naturally occurring host-pathogen interactions in vivo. We employed membrane shaving to identify peptide fragments cleaved from surface-expressed bacterial proteins and also detected proteins originating from the infected host. We optimised this technique for media-cultured Corynebacterium pseudotuberculosis, a sheep pathogen, revealing a set of 247 surface proteins. We then studied a natural host-pathogen interaction, by performing membrane shaving on C. pseudotuberculosis harvested directly from naturally infected sheep lymph nodes. Thirty-one bacterial surface proteins were identified including thirteen not identified in culture media, suggesting a different surface protein repertoire is expressed in this hostile environment. Forty-nine host proteins were identified including immune mediators and antimicrobial peptides such as cathelicidin. This novel application of proteolytic shaving has documented sets of host and pathogen proteins present at the bacterial surface in an infection of the native host.",
author = "Rees, {Megan A} and Oded Kleifeld and Crellin, {Paul K} and Ho, {Bosco Kwan-Chung} and Stinear, {Timothy P} and Smith, {Alexander Ian} and Coppel, {Ross L}",
year = "2015",
doi = "10.1021/pr5010086",
language = "English",
volume = "14",
pages = "120 -- 132",
journal = "Journal of Proteome Research",
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publisher = "American Chemical Society",
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Proteomic characterization of a natural host-pathogen interaction: repertoire of in vivo expressed bacterial and host surface-associated proteins. / Rees, Megan A; Kleifeld, Oded; Crellin, Paul K; Ho, Bosco Kwan-Chung; Stinear, Timothy P; Smith, Alexander Ian; Coppel, Ross L.

In: Journal of Proteome Research, Vol. 14, No. 1, 2015, p. 120 - 132.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Rees, Megan A

AU - Kleifeld, Oded

AU - Crellin, Paul K

AU - Ho, Bosco Kwan-Chung

AU - Stinear, Timothy P

AU - Smith, Alexander Ian

AU - Coppel, Ross L

PY - 2015

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N2 - Interactions between a host and a bacterial pathogen are mediated by cross-talk between molecules present on, or secreted by, pathogens and host binding-molecules. Identifying proteins involved at this interface would provide substantial insights into this interaction. While numerous studies have examined in vitro models of infection at the level of transcriptional change and proteomic profiling, there is virtually no information available on naturally occurring host-pathogen interactions in vivo. We employed membrane shaving to identify peptide fragments cleaved from surface-expressed bacterial proteins and also detected proteins originating from the infected host. We optimised this technique for media-cultured Corynebacterium pseudotuberculosis, a sheep pathogen, revealing a set of 247 surface proteins. We then studied a natural host-pathogen interaction, by performing membrane shaving on C. pseudotuberculosis harvested directly from naturally infected sheep lymph nodes. Thirty-one bacterial surface proteins were identified including thirteen not identified in culture media, suggesting a different surface protein repertoire is expressed in this hostile environment. Forty-nine host proteins were identified including immune mediators and antimicrobial peptides such as cathelicidin. This novel application of proteolytic shaving has documented sets of host and pathogen proteins present at the bacterial surface in an infection of the native host.

AB - Interactions between a host and a bacterial pathogen are mediated by cross-talk between molecules present on, or secreted by, pathogens and host binding-molecules. Identifying proteins involved at this interface would provide substantial insights into this interaction. While numerous studies have examined in vitro models of infection at the level of transcriptional change and proteomic profiling, there is virtually no information available on naturally occurring host-pathogen interactions in vivo. We employed membrane shaving to identify peptide fragments cleaved from surface-expressed bacterial proteins and also detected proteins originating from the infected host. We optimised this technique for media-cultured Corynebacterium pseudotuberculosis, a sheep pathogen, revealing a set of 247 surface proteins. We then studied a natural host-pathogen interaction, by performing membrane shaving on C. pseudotuberculosis harvested directly from naturally infected sheep lymph nodes. Thirty-one bacterial surface proteins were identified including thirteen not identified in culture media, suggesting a different surface protein repertoire is expressed in this hostile environment. Forty-nine host proteins were identified including immune mediators and antimicrobial peptides such as cathelicidin. This novel application of proteolytic shaving has documented sets of host and pathogen proteins present at the bacterial surface in an infection of the native host.

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