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Targeted comparative proteomics by liquid chromatography/matrix-assisted laser desorption/ionization triple-quadrupole mass spectrometry

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1002/rcm.2391
AuteurRechercher : 1; Rechercher : 1; Rechercher : 1
Affiliation
  1. Conseil national de recherches du Canada. Institut des biosciences marines du CNRC
FormatTexte, Article
Sujetliquid chromatography; proteomics; electrospray ionization; mass spectrometry; matrix-assisted laser desorption ionization; methods; amino acid sequence; metabolism; Candida albicans; pathogens; cattle; albumin; bovine; virulence; markers
Résumé
Here we report the first application of a matrix-assisted laser desorption/ionization (MALDI) triplequadrupole mass spectrometer for targeted proteomics. Employing an amine-specific isotopic labelling approach, the technique was validated using five randomly selected bovine serum albumin peptides differentially labelled at known ratios. An indirect benefit of the isotopic labelling technique is a significant enhancement of the a1 ion in tandem mass (MS/MS) spectra of all peptides studied. Therefore, the a1 ion was selected as the fragment ion for multiple reaction monitoring (MRM) in all cases, eliminating tedious method development and optimization. Accurate quantification was achieved with an average relative standard deviation (RSD) of 5% (n¼5) and a detection limit of 14 amol. The technique was then applied to validate an important virulence biomarker of the fungal pathogen Candida albicans, which was not accurately quantified using global proteomics experiment employing two-dimensional liquid chromatography/electrospray ionization tandem mass spectrometry (2D-LC/ESI)-MS/MS. Using LC/MALDI-MRM analysis of five tryptic peptides, the protein PHR1 was found to be upregulated in the hyphal (pathogenic) form of C. albicans by a factor of 7.7W0.8.
Date de publication
Conditions d’utilisation
  • Copyright © 2006 John Wiley & Sons, Ltd.
Dans
  • Rapid Communications in Mass Spectrometry 20, nº 5 (8 février 2006) : 904–910.
Langueanglais
Publications évaluées par des pairsOui
Numéro du CNRCNRCC 55833
1574
Numéro NPARC3538457
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Identificateur de l’enregistrement3ae13d5c-91d3-4709-86f8-c6bbdfe5a42b
Enregistrement créé2009-03-01
Enregistrement modifié2020-04-22
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