Rapid identification of pyoverdines of fluorescent Pseudomonas spp. by UHPLC-IM-MS.
Karoline RehmVera VollenweiderRolf KümmerliLaurent BiglerPublished in: Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine (2022)
Siderophores are iron-chelating molecules produced by bacteria and other microbes. They are involved with virulence in infections and play key roles in bacterial community assembly and as plant protectants due to their pathogen control properties. Although assays exist to screen whether newly isolated bacteria can produce siderophores, the chemical structures of many of these bio-active molecules remain unidentified due to the lack of rapid analytical procedures. An important group of siderophores are pyoverdines. They consist of a structurally diverse group of chromopeptides, whose amino acid sequence is characteristic for the fluorescent Pseudomonas species that secrets them. Although over 60 pyoverdine structures have been described so far, their characterization is cumbersome and several methods (isoelectrofocusing, iron uptake measurement, mass determination) are typically combined as ambiguous results are often achieved by a single method. Those additional experiments consume valuable time and resources and prevent high-throughput analysis. In this work, we present a new pyoverdine characterisation option by recording their collision cross sections (CCS) using trapped ion mobility spectrometry. This can be done simultaneously in combination with UHPLC and high-resolution MS resulting in a rapid identification of pyoverdines. The high specificity of CCS values is presented for 17 pyoverdines secreted by different Pseudomonas strains. The pyoverdine mass determination by full scan MS was supported by fragments obtained from broadband collision induced dissociation (bbCID). As iron contaminations in laboratories are not uncommon, CCS values of ferripyoverdines were also evaluated. Thereby, unusual and highly characteristic ion mobility patterns were obtained that are suitable as an alternative identification marker.
Keyphrases
- ms ms
- high resolution
- high throughput
- mass spectrometry
- solid phase extraction
- biofilm formation
- multiple sclerosis
- amino acid
- escherichia coli
- liquid chromatography
- tandem mass spectrometry
- quantum dots
- bioinformatics analysis
- loop mediated isothermal amplification
- pseudomonas aeruginosa
- molecularly imprinted
- computed tomography
- simultaneous determination
- iron deficiency
- staphylococcus aureus
- gas chromatography
- single cell
- high speed
- living cells
- ultra high performance liquid chromatography
- high resolution mass spectrometry
- high glucose
- magnetic resonance imaging
- label free
- oxidative stress
- sensitive detection
- endothelial cells
- single molecule
- cystic fibrosis
- stress induced