Characterisation of proteins extracted from the surface of Salmonella Typhimurium grown under SPI-2-inducing conditions by LC-ESI/MS/MS sequencing

Aileen E. Sherry, Neil F. Inglis, Andy Stevenson, Douglas J. Fraser-Pitt, Paul Everest, David George Emslie Smith, Mark Roberts

    Research output: Contribution to journalArticle

    Abstract

    Salmonella enterica has two pathogenicity islands encoding separate type three secretion systems (T3SS). Proteins secreted through these systems facilitate invasion and survival. After entry, Salmonella reside within a membrane bound vacuole, the Salmonella containing vacuole (SCV), where translocation of a second set of effectors by the Salmonella pathogenicity island 2 (SPI-2) T3SS is initiated. SPI-2 secretion in vitro can be induced by conditions that mimic the Salmonella containing vacuole. Utilising high-throughput mass spectrometry, we mapped the surface-attached proteome of S. Typhimurium SL1344 grown in vitro under SPI-2-inducing conditions and identified 108 proteins; using secretion signal prediction software, 43% of proteins identified contained a signal sequence. Of these proteins, 13 were known secreted effector proteins including SPI-2 effector proteins SseB, SseC, SseD, SseL, PipB2 and SteC, although surprisingly five were SPI-1 proteins, SipA, SipB, SipC, SipD and SopD, while 2 proteins SteA and SlrP are secreted by both T3SSs. This is the first in vitro study to demonstrate dual secretion of SPI-1 and SPI-2 proteins by S. Typhimurium and demonstrates the potential of high-throughput LC-ESI/MS/MS sequencing for the identification of novel proteins, providing a platform for subsequent comparative proteomic analysis, which should greatly assist understanding of the pathogenesis and inherent variation between serovars of Salmonella and ultimately help towards development of novel control strategies.

    Original languageEnglish
    Pages (from-to)361-370
    Number of pages10
    JournalProteomics
    Volume11
    Issue number3
    DOIs
    Publication statusPublished - Feb 2011

    Fingerprint

    Genomic Islands
    Salmonella typhimurium
    Salmonella
    Membrane Proteins
    Proteins
    Vacuoles
    Salmonella enterica
    Protein S
    Proteome
    Protein Sorting Signals
    Proteomics
    Mass Spectrometry
    Software
    Membranes

    Keywords

    • Bacterial Proteins
    • Blotting, Western
    • Chromatography, Liquid
    • Membrane Proteins
    • Salmonella Infections
    • Salmonella typhimurium
    • Signal Transduction
    • Spectrometry, Mass, Electrospray Ionization

    Cite this

    Sherry, Aileen E. ; Inglis, Neil F. ; Stevenson, Andy ; Fraser-Pitt, Douglas J. ; Everest, Paul ; Smith, David George Emslie ; Roberts, Mark. / Characterisation of proteins extracted from the surface of Salmonella Typhimurium grown under SPI-2-inducing conditions by LC-ESI/MS/MS sequencing. In: Proteomics. 2011 ; Vol. 11, No. 3. pp. 361-370.
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    abstract = "Salmonella enterica has two pathogenicity islands encoding separate type three secretion systems (T3SS). Proteins secreted through these systems facilitate invasion and survival. After entry, Salmonella reside within a membrane bound vacuole, the Salmonella containing vacuole (SCV), where translocation of a second set of effectors by the Salmonella pathogenicity island 2 (SPI-2) T3SS is initiated. SPI-2 secretion in vitro can be induced by conditions that mimic the Salmonella containing vacuole. Utilising high-throughput mass spectrometry, we mapped the surface-attached proteome of S. Typhimurium SL1344 grown in vitro under SPI-2-inducing conditions and identified 108 proteins; using secretion signal prediction software, 43{\%} of proteins identified contained a signal sequence. Of these proteins, 13 were known secreted effector proteins including SPI-2 effector proteins SseB, SseC, SseD, SseL, PipB2 and SteC, although surprisingly five were SPI-1 proteins, SipA, SipB, SipC, SipD and SopD, while 2 proteins SteA and SlrP are secreted by both T3SSs. This is the first in vitro study to demonstrate dual secretion of SPI-1 and SPI-2 proteins by S. Typhimurium and demonstrates the potential of high-throughput LC-ESI/MS/MS sequencing for the identification of novel proteins, providing a platform for subsequent comparative proteomic analysis, which should greatly assist understanding of the pathogenesis and inherent variation between serovars of Salmonella and ultimately help towards development of novel control strategies.",
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    note = "Copyright {\circledC} 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.",
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    Characterisation of proteins extracted from the surface of Salmonella Typhimurium grown under SPI-2-inducing conditions by LC-ESI/MS/MS sequencing. / Sherry, Aileen E.; Inglis, Neil F.; Stevenson, Andy; Fraser-Pitt, Douglas J.; Everest, Paul; Smith, David George Emslie; Roberts, Mark.

    In: Proteomics, Vol. 11, No. 3, 02.2011, p. 361-370.

    Research output: Contribution to journalArticle

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    AU - Sherry, Aileen E.

    AU - Inglis, Neil F.

    AU - Stevenson, Andy

    AU - Fraser-Pitt, Douglas J.

    AU - Everest, Paul

    AU - Smith, David George Emslie

    AU - Roberts, Mark

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    AB - Salmonella enterica has two pathogenicity islands encoding separate type three secretion systems (T3SS). Proteins secreted through these systems facilitate invasion and survival. After entry, Salmonella reside within a membrane bound vacuole, the Salmonella containing vacuole (SCV), where translocation of a second set of effectors by the Salmonella pathogenicity island 2 (SPI-2) T3SS is initiated. SPI-2 secretion in vitro can be induced by conditions that mimic the Salmonella containing vacuole. Utilising high-throughput mass spectrometry, we mapped the surface-attached proteome of S. Typhimurium SL1344 grown in vitro under SPI-2-inducing conditions and identified 108 proteins; using secretion signal prediction software, 43% of proteins identified contained a signal sequence. Of these proteins, 13 were known secreted effector proteins including SPI-2 effector proteins SseB, SseC, SseD, SseL, PipB2 and SteC, although surprisingly five were SPI-1 proteins, SipA, SipB, SipC, SipD and SopD, while 2 proteins SteA and SlrP are secreted by both T3SSs. This is the first in vitro study to demonstrate dual secretion of SPI-1 and SPI-2 proteins by S. Typhimurium and demonstrates the potential of high-throughput LC-ESI/MS/MS sequencing for the identification of novel proteins, providing a platform for subsequent comparative proteomic analysis, which should greatly assist understanding of the pathogenesis and inherent variation between serovars of Salmonella and ultimately help towards development of novel control strategies.

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    KW - Blotting, Western

    KW - Chromatography, Liquid

    KW - Membrane Proteins

    KW - Salmonella Infections

    KW - Salmonella typhimurium

    KW - Signal Transduction

    KW - Spectrometry, Mass, Electrospray Ionization

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