Analyses of the global secretome and whole cell proteome of a recent <em>Bordetella pertussis</em> epidemic strain — ASN Events
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  3. Analyses of the global secretome and whole cell proteome of a recent Bordetella pertussis epidemic strain

Analyses of the global secretome and whole cell proteome of a recent Bordetella pertussis epidemic strain (#428)

Laurence Luu 1 , Sophie Octavia 1 , Vitali Sintchenko 2 3 , Ling Zhong 4 , Mark Raftery 4 , Ruiting Lan 1
  1. School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, Australia
  2. Centre for Infectious Diseases and Microbiology–Public Health, Institute of Clinical Pathology and Medical Research, Westmead Hospital, New South Wales, Australia
  3. Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Sydney, New South Wales, Australia
  4. Mark Wainwright Analytical Centre, University of New South Wales, Sydney, New South Wales, Australia

Background: Bordetella pertussis is one of the most prevalent vaccine preventable diseases in Australia. The number of cases of B. pertussis has increased in the past decade even with high vaccination rates. Several factors have been hypothesised for the re-emergence of B. pertussis including waning immunity and pathogen adaptation. Despite decades of vaccination, our understanding of the biology of B. pertussis remains limited. We aimed to characterise the global secretome and whole cell proteome of L1423, a clinical isolate from the most recent Australian epidemic in 2008-2012. Methods: L1423 was grown in THIJS media at 37oC under shaking conditions for 12 hr. The supernatant was separated from the whole cell with centrifugation and then purified with a 3kDa filter. The whole cells were lysed with sonication and proteins were extracted and purified using acetone precipitation.  We then used LC-MS/MS to characterise the proteins found in the whole cell extract and supernatant. Results:  In the secretome, 140 proteins were identified with 74 proteins predicted to be secreted bioinformatically. Functional categories were assigned and the majority of proteins found were related to the cell surface with 31 proteins. Twenty three proteins were also associated with pathogenicity in the supernatant. Nine hundred and forty three proteins were found in the whole cell extract. The majority of these proteins were associated with cell surface and macromolecule synthesis or modification.  Forty four proteins were also found to be associated with virulence. Conclusion: This is the first study to characterise the global secretome and whole cell proteome of an Australian epidemic B. pertussis isolate.

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