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

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DOIResolve DOI: http://doi.org/10.1002/rcm.2391
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TypeArticle
Journal titleRapid Communications in Mass Spectrometry
ISSN09514198
Volume20
Issue5
Pages904910; # of pages: 7
Subjectliquid chromatography; proteomics; electrospray ionization; mass spectrometry; matrix-assisted laser desorption ionization; methods; amino acid sequence; metabolism; Candida albicans; pathogens; cattle; albumin; bovine; virulence; markers
AbstractHere 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.
Publication date
PublisherJohn Wiley & Sons, Ltd.
Copyright noticeCopyright © 2006 John Wiley & Sons, Ltd.
LanguageEnglish
AffiliationNRC Institute for Marine Biosciences; National Research Council Canada
Peer reviewedYes
NRC number55833
1574
NPARC number3538457
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Record identifier3ae13d5c-91d3-4709-86f8-c6bbdfe5a42b
Record created2009-03-01
Record modified2016-05-09
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