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

Sherry, A.E., Inglis, N.F., Stevenson, A., Fraser-Pitt, D.G.E., Everest, P. , Smith, D. and Roberts, M. (2011) Characterisation of proteins extracted from the surface of Salmonella Typhimurium grown under SPI-2-inducing conditions by LC-ESI/MS/MS sequencing. Proteomics, 11(3), pp. 361-370. (doi:10.1002/pmic.200900802)

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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

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Roberts, Professor Mark and Everest, Dr Paul and Stevenson, Mr Andrew and Smith, Professor David and Linn, Dr Aileen
Authors: Sherry, A.E., Inglis, N.F., Stevenson, A., Fraser-Pitt, D.G.E., Everest, P., Smith, D., and Roberts, M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Proteomics
ISSN:1615-9853
ISSN (Online):1615-9861
Published Online:03 January 2011

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
378891Secretomes of bacterial foodborne pathogens and functional assessment during interaction with intestinal epitheliumDavid SmithBiotechnology and Biological Sciences Research Council (BBSRC)BB/C508485/1Infection Immunity and Inflammation Life Sciences