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Microfluidics-based liquid chromatography/mass spectrometry multiple reaction monitoring approach for the relative quantification of Burkholderia cenocepacia secreted virulence factors

Sofie Depluverez (UGent) , Simon Daled (UGent) , Stijn De Waele (UGent) , Sören Planckaert (UGent) , Jolien Schoovaerts, Dieter Deforce (UGent) and Bart Devreese (UGent)
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Abstract
Rationale: Burkholderia cenocepacia is an opportunistic pathogen that is commonly isolated from patients with cystic fibrosis (CF). Quorum sensing has been suggested to play a role in the activity of type II and type VI secretion systems and the release of virulence factors. Apart from the classical acyl homoserine lactone quorum sensing, B. cenocepacia also uses the diffusible signal factor system (DSF). Quantitative information on the true impact of DSF molecules on the release of ZmpA and other virulence factors is lacking. Methods: Based on results of a label-free proteomics analysis addressing changes in the secretome in response to DSFs, a panel of peptides was selected to develop a microfluidics liquid chromatography/mass spectrometry (LC/MS) method implementing single reaction monitoring (SRM) to quantify B. cenocepacia virulence factors. Results: Increase in secretion of virulence factors upon treatment with BDSF was observed for ZmpA and Aida, but not for ZmpB. Type VI secretion system dependent Hcp1 and TecA were decreased. However, non-physiological amounts of BDSF were needed to provoke the effect. DSFs from P. aeruginosa and S. maltophilia were also affecting virulence factor secretion, but the effect was smaller than for the endogenous BDSF. Conclusions: Microfluidics-based SRM is a useful tool to quantitatively assess the impact of quorum sensing on the release of virulence factors by (opportunistic) pathogens.
Keywords
CYCLIC DI-GMP, CEPACIA COMPLEX, CYSTIC-FIBROSIS, PSEUDOMONAS-AERUGINOSA, STENOTROPHOMONAS-MALTOPHILIA, EPITHELIAL-CELLS, PHOSPHOLIPASE-A, SIGNAL, BACTERIA, ACID

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MLA
Depluverez, Sofie et al. “Microfluidics-based Liquid Chromatography/mass Spectrometry Multiple Reaction Monitoring Approach for the Relative Quantification of Burkholderia Cenocepacia Secreted Virulence Factors.” RAPID COMMUNICATIONS IN MASS SPECTROMETRY 32.6 (2018): 469–479. Print.
APA
Depluverez, S., Daled, S., De Waele, S., Planckaert, S., Schoovaerts, J., Deforce, D., & Devreese, B. (2018). Microfluidics-based liquid chromatography/mass spectrometry multiple reaction monitoring approach for the relative quantification of Burkholderia cenocepacia secreted virulence factors. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 32(6), 469–479.
Chicago author-date
Depluverez, Sofie, Simon Daled, Stijn De Waele, Sören Planckaert, Jolien Schoovaerts, Dieter Deforce, and Bart Devreese. 2018. “Microfluidics-based Liquid Chromatography/mass Spectrometry Multiple Reaction Monitoring Approach for the Relative Quantification of Burkholderia Cenocepacia Secreted Virulence Factors.” Rapid Communications in Mass Spectrometry 32 (6): 469–479.
Chicago author-date (all authors)
Depluverez, Sofie, Simon Daled, Stijn De Waele, Sören Planckaert, Jolien Schoovaerts, Dieter Deforce, and Bart Devreese. 2018. “Microfluidics-based Liquid Chromatography/mass Spectrometry Multiple Reaction Monitoring Approach for the Relative Quantification of Burkholderia Cenocepacia Secreted Virulence Factors.” Rapid Communications in Mass Spectrometry 32 (6): 469–479.
Vancouver
1.
Depluverez S, Daled S, De Waele S, Planckaert S, Schoovaerts J, Deforce D, et al. Microfluidics-based liquid chromatography/mass spectrometry multiple reaction monitoring approach for the relative quantification of Burkholderia cenocepacia secreted virulence factors. RAPID COMMUNICATIONS IN MASS SPECTROMETRY. 2018;32(6):469–79.
IEEE
[1]
S. Depluverez et al., “Microfluidics-based liquid chromatography/mass spectrometry multiple reaction monitoring approach for the relative quantification of Burkholderia cenocepacia secreted virulence factors,” RAPID COMMUNICATIONS IN MASS SPECTROMETRY, vol. 32, no. 6, pp. 469–479, 2018.
@article{8563992,
  abstract     = {Rationale: Burkholderia cenocepacia is an opportunistic pathogen that is commonly isolated from patients with cystic fibrosis (CF). Quorum sensing has been suggested to play a role in the activity of type II and type VI secretion systems and the release of virulence factors. Apart from the classical acyl homoserine lactone quorum sensing, B. cenocepacia also uses the diffusible signal factor system (DSF). Quantitative information on the true impact of DSF molecules on the release of ZmpA and other virulence factors is lacking. 
Methods: Based on results of a label-free proteomics analysis addressing changes in the secretome in response to DSFs, a panel of peptides was selected to develop a microfluidics liquid chromatography/mass spectrometry (LC/MS) method implementing single reaction monitoring (SRM) to quantify B. cenocepacia virulence factors. 
Results: Increase in secretion of virulence factors upon treatment with BDSF was observed for ZmpA and Aida, but not for ZmpB. Type VI secretion system dependent Hcp1 and TecA were decreased. However, non-physiological amounts of BDSF were needed to provoke the effect. DSFs from P. aeruginosa and S. maltophilia were also affecting virulence factor secretion, but the effect was smaller than for the endogenous BDSF. 
Conclusions: Microfluidics-based SRM is a useful tool to quantitatively assess the impact of quorum sensing on the release of virulence factors by (opportunistic) pathogens.},
  author       = {Depluverez, Sofie and Daled, Simon and De Waele, Stijn and Planckaert, Sören and Schoovaerts, Jolien and Deforce, Dieter and Devreese, Bart},
  issn         = {0951-4198},
  journal      = {RAPID COMMUNICATIONS IN MASS SPECTROMETRY},
  keywords     = {CYCLIC DI-GMP,CEPACIA COMPLEX,CYSTIC-FIBROSIS,PSEUDOMONAS-AERUGINOSA,STENOTROPHOMONAS-MALTOPHILIA,EPITHELIAL-CELLS,PHOSPHOLIPASE-A,SIGNAL,BACTERIA,ACID},
  language     = {eng},
  number       = {6},
  pages        = {469--479},
  title        = {Microfluidics-based liquid chromatography/mass spectrometry multiple reaction monitoring approach for the relative quantification of Burkholderia cenocepacia secreted virulence factors},
  url          = {http://dx.doi.org/10.1002/rcm.8059},
  volume       = {32},
  year         = {2018},
}

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