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

Abstract

Burkholderia cenocepacia is an opportunistic pathogen that is commonly isolated from patients with cystic fibrosis (CF). Several virulence factors have been identified, including extracellular enzymes that are secreted by type II and type VI secretion systems. The activity of these secretion systems is modulated by quorum sensing. Apart from the classical acylhomoserine lactone quorum sensing, B. cenocepacia also uses the diffusible signal factor system (DSF) i.e. 2-undecenoic acid derivatives that are recognized by specific receptors resulting in changes in biofilm formation, motility and virulence. However, quantitative information on alterations in the actual production and release of virulence factors upon exposure to DSF is lacking. We here describe an approach implementing microfluidics based chromatography combined with single reaction monitoring to quantify protein virulence factors in the secretome of B. cenocepacia.

Microfluidics for Peptidomics, Proteomics, and Cell Analysis

“…LC MF chips need to be coupled with MS detection to obtain relevant properties of the separated analytes, especially peptides and proteins. Depluverez et al [ 57 ] used the LC chip in conjunction with multiple reaction monitoring (MRM) MS detection to measure protein virulence factors of Burkholderia cenocepacia, an opportunistic pathogen commonly isolated from cystic fibrosis (CF) patients. Several virulence factors have been described, including extracellular enzymes secreted by secretion systems of the II and VI types.…”

“…ESI and MALDI are the two most common methods for ionizing large (bio)molecules, especially polypeptides and proteins. Therefore, recent developments at the interface between MF microchips and mass spectrometers have focused on these two ionization methods [ 57 , 83 , 84 ].…”

<p>Quorum Quenching: A Potential Target for Antipseudomonal Therapy</p>

“…Subsequently, other DSF family signals were also described in Burkholderia cenocepacia and P. aeruginosa, which can synthesize cis-2-dodecenoic acid (BDSF) and cis-2-decenoic acid (PDSF), respectively. 17,21 There is a significant correlation among QS and the pathogenic factors production, motility, plasmid transfer, antibiotic production, and biofilm formation. Accordingly, the QS system facilitates the population to live and multiply in a better environment with effective intercellular communication, since it can contribute to several behaviors that enable bacteria to resist antibacterial compounds or antibiotics like biofilm development.…”